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Differential Induction of Colitis and Gastritis in HLA-B27 Transgenic Rats Selectively Colonized with Bacteroides vulgatus or Escherichia coli  

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Resident bacteria play an important role in initiating and perpetuating gastrointestinal inflammation. We previously demonstrated that six commensal bacteria including Bacteroides vulgatus caused more aggressive colitis and gastritis in HLA-B27 transgenic rats than did the other five bacteria without B. vulgatus. This study compared the degree of gastrointestinal inflammation in gnotobiotic HLA-B27 transgenic rats monoassociated with either B. vulgatus or Escherichia coli. Gnotobiotic transge...

Rath, Heiko C.; Wilson, Kenneth H.; Sartor, R. Balfour

1999-01-01

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HLA-B27 heavy chain homodimers are expressed in HLA-B27 transgenic rodent models of spondyloarthritis and are ligands for paired Ig-like receptors.  

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HLA-B27 transgenic rats and strains of HLA-B27-transgenic beta(2)-microglobulin (beta(2)m)-deficient mice develop a multisystem inflammatory disease affecting the joints, skin, and bowel with strong similarity to human spondyloarthritis. We show that HLA-B27 transgenic mice and rats express HC10-reactive, beta(2)m-free HLA-B27 homodimers (B27(2)) and multimers, both intracellularly and at the cell surface of leukocytes, including rat dendritic cells. Fluorescent-labeled tetrameric complexes o...

Kollnberger, S.; Bird, La; Roddis, M.; Hacquard-bouder, C.; Kubagawa, H.; Bodmer, Hc; Breban, M.; Mcmichael, Aj; Bowness, P.

2004-01-01

3

The Specificity of Peptides Bound to Human Histocompatibility Leukocyte Antigen (HLA)-B27 Influences the Prevalence of Arthritis in HLA-B27 Transgenic Rats  

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Human histocompatibility leukocyte antigen B27 is highly associated with the rheumatic diseases termed spondyloarthropathies, but the mechanism is not known. B27 transgenic rats develop a spontaneous disease resembling the human spondyloarthropathies that includes arthritis and colitis. To investigate whether this disease requires the binding of specific peptides to B27, we made a minigene construct in which a peptide from influenza nucleoprotein, NP383-391 (SRYWAIRTR), which binds B27 wit...

Zhou, Ming; Sayad, Alain; Simmons, William A.; Jones, Richard C.; Maika, Shanna D.; Satumtira, Nimman; Dorris, Martha L.; Gaskell, Simon J.; Bordoli, Robert S.; Balfour Sartor, R.; Slaughter, Clive A.; Richardson, James A.; Hammer, Robert E.; Taurog, Joel D.

1998-01-01

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Spontaneous inflammatory arthritis in HLA-B27 transgenic mice lacking beta 2-microglobulin: a model of human spondyloarthropathies  

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Human class I major histocompatibility complex allele HLA-B27 is associated with a group of human diseases called "spondyloarthropathies." Studies on transgenic rats expressing HLA-B27 and human beta 2-microglobulin have confirmed the role of HLA-B27 in disease pathogenesis. Here we report spontaneous inflammatory arthritis in HLA-B27 transgenic mice lacking beta 2-microglobulin (B27+ beta 2m-/- ). In the absence of beta 2-microglobulin, B27+ beta 2m-/- animals do not express the HLA-B27 tran...

1995-01-01

5

The germfree state prevents development of gut and joint inflammatory disease in HLA-B27 transgenic rats  

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A number of inflammatory disease states occur with greatly increased frequency in individuals inheriting the human major histocompatibility complex class I allele HLA-B27. In a minority of cases, namely those with B27-associated reactive arthritis, there is good evidence that the disease state is triggered by infection with an enteric or genitourinary bacterial pathogen. For the majority of B27-associated disease, no definite pathogenetic role for bacteria has been established. However, in th...

1994-01-01

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Effects of Feeding a Probiotic Preparation (SIM) Containing Inulin on the Severity of Colitis and on the Composition of the Intestinal Microflora in HLA-B27 Transgenic Rats  

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An overly aggressive immune response to the intestinal microflora in a genetically susceptible host background has been implicated in the pathogenesis of inflammatory bowel diseases. We measured the impact of a probiotic preparation (SIM) containing inulin on the severity of colitis and on intestinal microflora profiles of HLA-B27-?2-microglobulin transgenic (TG) rats. SIM is a mixture of lactobacilli, bifidobacteria, and inulin. Two-month-old TG rats received either SIM or water. Control TG...

Schultz, M.; Munro, K.; Tannock, G. W.; Melchner, I.; Go?ttl, C.; Schwietz, H.; Scho?lmerich, J.; Rath, H. C.

2004-01-01

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Chemically Defined Diet Alters the Protective Properties of Fructo-Oligosaccharides and Isomalto-Oligosaccharides in HLA-B27 Transgenic Rats  

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Non-digestible oligosaccharides (NDO) were shown to reduce inflammation in experimental colitis, but it remains unclear whether microbiota changes mediate their colitis-modulating effects. This study assessed intestinal microbiota and intestinal inflammation after feeding chemically defined AIN-76A or rat chow diets, with or without supplementation with 8 g/kg body weight of fructo-oligosaccharides (FOS) or isomalto-oligosaccharides (IMO). The study used HLA-B27 transgenic rats, a validated model of inflammatory bowel disease (IBD), in a factorial design with 6 treatment groups. Intestinal inflammation and intestinal microbiota were analysed after 12 weeks of treatment. FOS and IMO reduced colitis in animals fed rat chow, but exhibited no anti-inflammatory effect when added to AIN-76A diets. Both NDO induced specific but divergent microbiota changes. Bifidobacteria and Enterobacteriaceae were stimulated by FOS, whereas copy numbers of Clostridium cluster IV were decreased. In addition, higher concentrations of total short-chain fatty acids (SCFA) were observed in cecal contents of rats on rat chow compared to the chemically defined diet. AIN-76A increased the relative proportions of propionate, iso-butyrate, valerate and iso-valerate irrespective of the oligosaccharide treatment. The SCFA composition, particularly the relative concentration of iso-butyrate, valerate and iso-valerate, was associated (P?0.004 and r?0.4) with increased colitis and IL-1 ? concentration of the cecal mucosa. This study demonstrated that the protective effects of fibres on colitis development depend on the diet. Although diets modified specific cecal microbiota, our study indicates that these changes were not associated with colitis reduction. Intestinal inflammation was positively correlated to protein fermentation and negatively correlated with carbohydrate fermentation in the large intestine. PMID:25369019

Valcheva, Rosica; Gänzle, Michael G.; Dieleman, Levinus A.

2014-01-01

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Fully functional HLA B27-restricted CD4+ as well as CD8+ T cell responses in TCR transgenic mice.  

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The strong association of HLA B27 with spondyloarthropathies contrasts strikingly with most autoimmune diseases, which are HLA class II associated and thought to be mediated by CD4+ T lymphocytes. By introducing a human-derived HLA B27-restricted TCR into HLA B27 transgenic mice, we have obtained a functional TCR transgenic model, GRb, dependent on HLA B27 for response. Surprisingly, HLA B27 supported CD4+ as well as CD8+ T cell responses in vivo and in vitro. Further, HLA B27-restricted CD4+...

Roddis, M.; Carter, Rw; Sun, My; Weissensteiner, T.; Mcmichael, Aj; Bowness, P.; Bodmer, Hc

2004-01-01

9

Sharing of an HLA-B27-restricted H-Y antigen between rat and mouse.  

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The purpose of this work was twofold: 1) to learn whether rats transgenic for HLA-B27 and the human beta 2-microglobulin gene HB2M can mount B27-restricted cytolytic T lymphocyte (CTL) responses to the male H-Y antigen, and 2) to learn whether such CTLs would recognize both rat and mouse H-Y in the context of HLA-B27. Female rats of the B27/HB2M transgenic line 21-4L were primed in vivo with cells from males of the same line. CTL effectors were generated from lymph node cells of these females following culture with irradiated antigen-presenting cells from either male 21-4L rats or male mice of the B27/HB2M transgenic 56-3 line. The CTLs showed male-specific, B27-specific lysis of both rat and mouse targets. Lysis of B27 targets was inhibitable by monoclonal antibodies specific for B27 or rat CD8. Specific lysis of male B27 rat and mouse targets was inhibitable equally by either rat or mouse male B27 cold targets, but not significantly by female or nontransgenic cold targets. The B27-restricted CTLs neither recognized nor were inhibited by B27+ or B27- male or female human targets. These results demonstrate that CD8+, B27-restricted, anti-H-Y CTLs recognize an evolutionarily conserved H-Y peptide antigen in both rats and mice. In addition, they establish the transgenic rat as a model system for examining the T-cell response to antigen presented by class I HLA molecules. PMID:8344721

Simmons, W A; Taurog, J D; Hammer, R E; Breban, M

1993-01-01

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Expression of HLA-B27 in transgenic mice is dependent on the mouse H-2D genes  

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HLA-B27 transgenic mice in the context of various H-2 haplotypes were produced. A high expression of the HLA-B27 antigen was observed in mice homozygous for H-2b, H-2f, H-2s, H-2p, H-2r, and H-2k haplotypes. Mice of the H-2v haplotype expressed HLA-B27 at an intermediate level. Expression of HLA-B27 was minimal in mice of the H-2q and H-2d haplotypes. This was observed both on the B10 background and in DBA/2 or BALB/c mice. Only minimal expression of HLA-B27 could be detected in B10.PL (KuDd)...

1990-01-01

11

Altered bone material properties in HLA-B27 rats include reduced mineral to matrix ratio and altered collagen cross-links.  

Science.gov (United States)

Spondyloarthropathy and inflammatory bowel disease (IBD), which includes ulcerative colitis and Crohn's disease, are often associated with severe osteopenia/osteoporosis in both children and adults. HLA-B27 transgenic rats present a phenotype that includes severe colitis and severely accelerated alveolar bone loss. The purpose of this study was to evaluate long bone density status, systemic bone metabolic markers, and intrinsic bone material properties in HLA-B27 transgenic (TG) rats, and compare them with those of age- and sex-matched wild-type (WT) animals. The results indicate that in the HLA-B27 rat, an animal susceptible to both alveolar bone loss (ABL) and long bone osteopenia, there is a statistically significant negative correlation between ABL and long bone bone mineral density (BMD), as well as mineral/matrix ratio at active bone-forming trabecular surfaces. The TG animals had a lower mineral/matrix ratio and higher relative proteoglycan and advanced glycation end product (?-N-Carboxymethyl-L-lysine) content and pyridinoline/divalent collagen cross-link ratio compared with WT. These results may provide better understanding of the interrelationship between osteoporosis and oral bone loss, the underlying causes of the inferior bone strength in the HLA-B27 transgenic animals, and could prove to be a useful model in the elucidation of the pathophysiology of spondyloarthropathy and IBD-associated osteopenia/osteoporosis and in the evaluation of pharmacological intervention(s) against such conditions. PMID:24771481

Gamsjaeger, Sonja; Srivastava, Apurva K; Wergedal, Jon E; Zwerina, Jochen; Klaushofer, Klaus; Paschalis, Eleftherios P; Tatakis, Dimitris N

2014-11-01

12

HLA-B27 heavy chains contribute to spontaneous inflammatory disease in B27/human beta2-microglobulin (beta2m) double transgenic mice with disrupted mouse beta2m.  

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MHC class I allele, HLA-B27, is strongly associated with a group of human diseases called spondyloarthropathies. Some of these diseases have an onset after an enteric or genitourinary infection. In the present study, we describe spontaneous disease in HLA-B27 transgenic mice where endogenous beta2-microglobulin (beta2m) gene was replaced with transgenic human beta2m gene. These mice showed cell surface expression of HLA-B27 similar to that of human peripheral blood mononuclear cells. In addit...

Khare, S. D.; Hansen, J.; Luthra, H. S.; David, C. S.

1996-01-01

13

HLA-B27 antigen  

Science.gov (United States)

HLA-B27 is a blood test to look for a protein that is found on the surface of ... The protein is called human leukocyte antigen B27 (HLA-B27). Human leukocyte antigens (HLAs) are proteins that help ...

14

Identification of HLA-B27-restricted peptides from the Chlamydia trachomatis proteome with possible relevance to HLA-B27-associated diseases.  

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The association of HLA-B27 with ankylosing spondylitis and reactive arthritis is the strongest one known between an MHC class I Ag and a disease. We have searched the proteome of the bacterium Chlamydia trachomatis for HLA-B27 binding peptides that are stimulatory for CD8(+) cells both in a model of HLA-B27 transgenic mice and in patients. This was done by combining two biomathematical computer programs, the first of which predicts HLA-B27 peptide binding epitopes, and the second the probabil...

Kuon, W.; Holzhu?tter, Hg; Appel, H.; Grolms, M.; Kollnberger, S.; Traeder, A.; Henklein, P.; Weiss, E.; Thiel, A.; Lauster, R.; Bowness, P.; Radbruch, A.; Kloetzel, Pm; Sieper, J.

2001-01-01

15

Expression of aberrant HLA-B27 molecules is dependent on B27 dosage and peptide supply.  

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OBJECTIVES: Cellular expression of non-classical forms of human leukocyte antigen (HLA)-B27 (NC-B27) may be involved in spondyloarthritis (SpA) pathogenesis. We used a novel B27-specific monoclonal antibody, HD6, to ask if B27 transgenic (TG) rat splenocytes express these NC-B27 molecules. We also investigated whether B27-binding peptides could affect the expression and functional immune recognition of HD6-reactive B27 molecules. METHODS: Splenocytes from B27-TG, B7-TG and non-transgenic rats...

Mchugh, K.; Rysnik, O.; Kollnberger, S.; Shaw, J.; Utriainen, L.; Al-mossawi, Mh; Payeli, S.; Marroquin, O.; Milling, S.; Renner, C.; Bowness, P.

2014-01-01

16

Heart conduction disturbance: an HLA-B27 associated disease.  

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In recent studies from Sweden an increased prevalence of HLA-B27 associated diseases and of HLA-B27 was found in an unselected group of men with permanently implanted pacemakers and with a heart block. Furthermore, a significantly increased prevalence of HLA-B27 was found in men with a pacemaker who had no clinical or radiological signs of HLA-B27 associated disease. To obtain more insight into the association between HLA-B27 and heart block, and the possible role of HLA-B27 in causing this b...

Peeters, A. J.; Ten Wolde, S.; Sedney, M. I.; Vries, R. R.; Dijkmans, B. A.

1991-01-01

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HLA B27 y las espondilartropatías seronegativas  

Directory of Open Access Journals (Sweden)

Full Text Available Se realizó el tipaje serológico para el antígeno HLA B27 a 19 pacientes con espondilartropatías seronegativas para conocer su relación y, de ellos, 6 resultaron positivos; 94 individuos sanos conformaron el grupo control y en 4 se encontró el antígeno. Los resultados expuestos sugieren la presencia de genes adicionales al B27 en los pacientes con este grupo de enfermedades.The serologic typing of HLA B27 antigen was perfomed in 19 patients presenting with seronegative spondyloarthropathies in order to know its relationship. Of them 6 patients were found to be positive; 94 healthy subjects were inclkuded in the control group and 4 presented with the antigen. Results reported suggest the presence of additional genes to B27 in patients presenting with this group of diseases.

Modesto González Cortiñas

1997-04-01

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HLA B27 y las espondilartropatías seronegativas  

Scientific Electronic Library Online (English)

Full Text Available SciELO Cuba | Language: Spanish Abstract in spanish Se realizó el tipaje serológico para el antígeno HLA B27 a 19 pacientes con espondilartropatías seronegativas para conocer su relación y, de ellos, 6 resultaron positivos; 94 individuos sanos conformaron el grupo control y en 4 se encontró el antígeno. Los resultados expuestos sugieren la presencia [...] de genes adicionales al B27 en los pacientes con este grupo de enfermedades. Abstract in english The serologic typing of HLA B27 antigen was perfomed in 19 patients presenting with seronegative spondyloarthropathies in order to know its relationship. Of them 6 patients were found to be positive; 94 healthy subjects were inclkuded in the control group and 4 presented with the antigen. Results re [...] ported suggest the presence of additional genes to B27 in patients presenting with this group of diseases.

Modesto, González Cortiñas; Lourdes, Faurés Vergara; Ricardo, Rodríguez Viera; Jesús, Gómez Arbesú.

1997-04-01

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HLA-B27 subtypes among the Chukotka native groups  

International Nuclear Information System (INIS)

The purpose of this study was to assess the relative frequency of the known HLA-B27 subtypes in HLA-B27 positive Chukotka natives, which have higher frequencies of HLA-B27 (to 40%) and spondylarthropathies (to 2%) than the Russian Caucasian population. Using oligotyping of the polymerase-chain reaction amplified second and third exons of the HLA-B27 gene in 86 DNA samples from HLA-B27 positive individuals were successfully typed. All had HLA-B*2705, including 4 patients with Reiter's syndrome and 5 with ankylosing spondyloarthritis, except one Eskimo who had HLA-B*2702. None had HLA-B*2704, a frequent subtype in Orientals. With respect to HLA-B27 subtypes the indigenous populations from the eastern part of the Chukotka Peninsula are genetically more closely related to Caucasians than to Orientals. (author). 18 refs, 1 fig., 2 tabs

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HLA-B27 and an electrocardiographic peculiarity  

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INTRODUCTION: An increased cardiovascular mortality has been described in patients with spondyloarthropathies due to HLA-B27. Numerous cardiovascular afflictions are currently known to be associated with HLA-B27. These include aortic root dilation, aortic regurgitation, mitral regurgitation, myocarditis, heart failure, pericarditis, pericardial effusion, atrioventricular conduction block and more recently, the presence of J-waves. MATERIALS AND METHODS: 48 HLA-B27 positive patient...

Ker, James

2011-01-01

 
 
 
 
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HLA-B27 typing in the categorisation of uveitis in a HLA-B27 rich population  

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AIMS—To determine whether HLA-B27 typing helps the clinician in the diagnostic examination of uveitis in a HLA-B27 rich population and also whether the clinical picture of HLA-B27 positive unilateral acute or recurrent anterior uveitis (AAU) is distinguishable from the idiopathic negative form.?METHODS—During a 3 year period 220 consecutive patients with undetermined uveitis at onset were examined in the Helsinki University Eye Clinic. HLA-B27 antigen was tested for 85% of the patient...

Huhtinen, M.; Karma, A.

2000-01-01

22

Isolated HLA-B27 associated Achilles tendinitis.  

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The case of a 37 year old man with a longstanding HLA-B27 associated bilateral Achilles tendinitis without seronegative spondyloarthropathy is reported. This case suggests that heel enthesopathy may for a long time be the only clinical manifestation of the HLA-B27 associated disease process.

Olivieri, I.; Gemignani, G.; Gherardi, S.; Grassi, L.; Ciompi, M. L.

1987-01-01

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Bartonella henselae associated uveitis and HLA-B27  

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AIM—To investigate the frequency of HLA-B27 in patients with presumed Bartonella henselae associated uveitis and to describe the clinical characteristics of HLA-B27 positive patients with uveitis and presumed ocular bartonellosis (POB).?METHODS—The diagnosis of POB was considered in 19 patients with unexplained uveitis (except for the HLA-B27 association) and high positive IgG (titre ?1:900) and/or IgM (titre ?1:250) antibodies against B henselae. In addition to B henselae serology...

Kerkhoff, F. T.; Rothova, A.

2000-01-01

24

Multiple joint tuberculosis presenting as HLA-B27 disease  

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A case of multifocal osteoarticular tuberculosis in a young Caucasian male is presented. The diagnostic difficulty, compounded by slow progression of the disorder and the presence of the tissue antigen HLA-B27, is discussed.

Hopkins, G. O.

1983-01-01

25

Coexisting HLA-B27 positive spondyloarthritis and polyarteritis nodosa.  

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A 38 year old woman presented with widespread polyarteritis nodosa a few years after the onset of HLA-B27 positive spondyloarthritis. The concomitant coexistence of these two disorders suggests a possible association in this genetically susceptible subject.

Sattar, M. A.

1992-01-01

26

Jekyll and Hyde: the transformation of HLA-B27.  

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HLA-B27 can adopt a homodimeric form. In spondyloarthritis, such aberrant expression of B27 may override a tissue-specific inhibition of major histocompatibility complex (MHC) class I-dependent events, leading to inflammation and fibrosis.

Edwards, Jc; Bowness, P.; Archer, Jr

2000-01-01

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Panuveíte em artrite indiferenciada HLA-B27 positiva Panuveitis in HLA-B27 positive undifferentiated arthritis  

Directory of Open Access Journals (Sweden)

Full Text Available Entre os vários tipos de inflamação ocular associados às doenças reumatológicas, a uveíte anterior é particularmente comum nas espondiloartropatias, em especial quando associada à presença do genótipo HLA-B27. Relatou-se o caso de um paciente com artrite indiferenciada HLA-B27 positivo, complicado com panuveíte e vasculite da retina, refratária ao tratamento imunossupressor tradicional, que obteve boa resposta clínica ao uso de anti-TNF-alfa.Among the several types of ocular inflammation associated to the rheumatic diseases, anterior uveitis is particularly common in the spondyloarthropathies, especially when associated to the presence of the HLA-B27 genotype. We report the case of HLA-B27 positive patient with undifferentiated arthritis, complicated with panuveitis and retinal vasculitis, that was refractory to the traditional imunossupressive treatment, and had a good clinical response with anti-TNF-alpha therapy.

Mário Sérgio Ferreira Santos

2008-10-01

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Panuveíte em artrite indiferenciada HLA-B27 positiva / Panuveitis in HLA-B27 positive undifferentiated arthritis  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese Entre os vários tipos de inflamação ocular associados às doenças reumatológicas, a uveíte anterior é particularmente comum nas espondiloartropatias, em especial quando associada à presença do genótipo HLA-B27. Relatou-se o caso de um paciente com artrite indiferenciada HLA-B27 positivo, complicado c [...] om panuveíte e vasculite da retina, refratária ao tratamento imunossupressor tradicional, que obteve boa resposta clínica ao uso de anti-TNF-alfa. Abstract in english Among the several types of ocular inflammation associated to the rheumatic diseases, anterior uveitis is particularly common in the spondyloarthropathies, especially when associated to the presence of the HLA-B27 genotype. We report the case of HLA-B27 positive patient with undifferentiated arthriti [...] s, complicated with panuveitis and retinal vasculitis, that was refractory to the traditional imunossupressive treatment, and had a good clinical response with anti-TNF-alpha therapy.

Mário Sérgio Ferreira, Santos; Vitor, Cortizo; Cícero Ricardo Torres da, Costa; Ronnielly Melo, Tavares; Ricardo Eric Barros, Lopes.

2008-10-01

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Panuveíte em artrite indiferenciada HLA-B27 positiva Panuveitis in HLA-B27 positive undifferentiated arthritis  

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Entre os vários tipos de inflamação ocular associados às doenças reumatológicas, a uveíte anterior é particularmente comum nas espondiloartropatias, em especial quando associada à presença do genótipo HLA-B27. Relatou-se o caso de um paciente com artrite indiferenciada HLA-B27 positivo, complicado com panuveíte e vasculite da retina, refratária ao tratamento imunossupressor tradicional, que obteve boa resposta clínica ao uso de anti-TNF-alfa.Among the several types of ocular...

Mário Sérgio Ferreira Santos; Vitor Cortizo; Cícero Ricardo Torres da Costa; Ronnielly Melo Tavares; Ricardo Eric Barros Lopes

2008-01-01

30

Invasion and persistence of Salmonella in human fibroblasts positive or negative for endogenous HLA B27  

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OBJECTIVE—Analysis of the interaction of enteropathogenic bacteria with HLA B27 transfected murine fibroblasts showed a specific influence of HLA B27 on microbial invasiveness. This possible novel mechanism for the action of HLA B27 should be verified by using endogenous HLA B27 positive and negative human fibroblasts as a model for the direct interaction of arthritogenic bacteria and host cells.?METHODS—Fibroblasts were obtained from healthy donors positive or negative for HLA B27; cul...

Huppertz, H.; Heesemann, J.

1997-01-01

31

HLA-B27, but not HLA-B7, immunodominance to influenza is ERAP dependent.  

Science.gov (United States)

Endoplasmic reticulum-associated aminopeptidase-1 (ERAP1) plays a critical role in the processing of peptides prior to binding to MHC class I molecules. In this article, we show for the first time, to our knowledge, that the HLA-B27 immunodominant influenza nucleoprotein (NP) 383-391 epitope is made as an N-terminally extended 14-mer before it is trimmed by ERAP. In the absence of ERAP, there is a significant reduction in the CTL response to the B27/NP383-391 epitope in influenza A (flu)-infected B27/ERAP(-/-) mice. With the use of tetramer staining, the number of naive CD8(+) T cells expressing TCR V?8.1 in B27/ERAP(-/-) transgenic mice is significantly lower than that seen in B27/ERAP(+/+) mice. HLA-B27 surface expression in naive and flu-infected B27/ERAP(-/-) mice is also lower than the expression seen for the same allele in naive and flu-infected B27/ERAP(+/+) mice. In contrast, surface expression of HLA-B7 was unaffected by the absence of ERAP in B7/ERAP(-/-) transgenic mice. The B7-restricted NP418-426 CTL response in flu-infected B7/ERAP(-/-) and B7/ERAP(+/+) mice was also similar. These results provide, to our knowledge, the first in vivo demonstration of ERAP functionally influencing host immune response in an HLA allele-specific manner. This principle has relevance to diseases such as ankylosing spondylitis, in which HLA-B27 and ERAP jointly contribute to disease predisposition. PMID:24835397

Akram, Ali; Lin, Aifeng; Gracey, Eric; Streutker, Catherine J; Inman, Robert D

2014-06-15

32

HLA-B27 frequency in a group of patients with psoriatic arthritis Freqüência de HLA-B27 em uma amostra de pacientes com artrite psoriática  

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BACKGROUND: HLA-B27 is associated with spondyloarthritis, a group of diseases that includes psoriatic arthritis. OBJECTIVES: To describe the HLA-B27 frequency in a group of Brazilian patients with psoriatic arthritis and correlate its presence or absence with their clinical manifestations. METHODS: Cross-sectional study with 44 psoriatic arthritis patients of a Rheumatology clinic. Demographic and social data were recorded, as were skin and joints clinical examination. HLA-B27 was tested. All...

Danilo Garcia Ruiz; Mário Newton Leitão de Azevedo; Omar Lupi

2012-01-01

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Acute anterior uveitis, ankylosing spondylitis, back pain, and HLA-B27.  

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One hundred and sixty-nine patients with acute anterior uveitis were studied for the presence of HLA-B27 tissue type, radiological evidence of ankylosing spondylitis, and a history of back pain. 60% were male; 45% were HLA-B27+. The male:female ratio in the HLA-B27+ group was the same as in the whole group. 24% had radiological evidence of ankylosing spondylitis, and, of these, 83% were HLA-B27+ while 17% were HLA-B27-. There was a definite correlation between the severity of the ankylosing s...

Beckingsale, A. B.; Davies, J.; Gibson, J. M.; Rosenthal, A. R.

1984-01-01

34

HLA-B*27 typing by sequence specific amplification without DNA extraction.  

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HLA-B27 appears to play a direct role in the pathogenesis of ankylosing spondylitis and almost all patients with this disease have HLA-B27. Therefore, a diagnosis of ankylosing spondylitis can virtually be excluded in the absence of HLA-B27. Many techniques have been used for HLA-B*27 typing. Of these, molecular methods are the most sensitive and specific but require extracted DNA as the testing material. A technique where HLA-B*27 is amplified directly from whole blood using sequence specifi...

Sayer, D. C.; Cassell, H. S.; Christiansen, F. T.

1999-01-01

35

Role of HLA B27 in diagnosis of seronegative spondyloarthropathies.  

Science.gov (United States)

Seronegative Spondyloarthropathies (SSA) is a very common problem in our area. The main aim of present study was (1) to find the HLA B27 positivity in patients presenting with sacroileitis (2) to see the correlation of B27 positivity on haematological, radiological and extra articular manifestations. Total 110 patients of SSA were studied between July 2004 to June 2005. Routine haematological and immunological test were done by standard method. Total positivity of B27 in SSA was 43.63%, HLA B27 positivity was higher in children (68.75%). Sex wise analysis of B27 positive cases showed that 81.81% B27 positive patients were males. In HLA B27 positive cases lower spine, hip, sacroiliac, shoulder and knee joints were more involved (77.08%, 79.16%, 79.16%, 37.50% and 50.00% respectively). Urinary tract infection (UTI), diarrhoea and constipation were more common in B27 positive cases. Leukocytosis of neutrophilic type (33.33%), raised ESR (77.55%)., CRP positivity (63.63%) and anaemia (65.00%) were seen more frequently in B27 positive cases. In bilateral sacroiliitis diagnosed by X-ray, only 69.23% patient were B27 positive. Our study concludes that HLA B 27 positivity is higher in SSA seen in childhood and in young adult males. B27 positive patients have more severe disease and systemic manifestation Hence, male patients specially young adolescent or young adults with sacroileitis must be subjected for B27 typing. PMID:18306603

Sonkar, Gyanendra Kumar; Usha

2007-10-01

36

Arthrite rhumatoïde juvénile et HLA-B27.  

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Forty children with juvenile rheumatoid arthritis were studied to determine the frequency of the histocompatibility antigen HLA [human leukocyte antigen)-B27 in this disease and to characterize the arthropathy associated with this antigen. HLA-B27 was detected in four patients (10%). Its presence was associated in a statistically significant manner with sacroiliitis demonstrated radiologically and with a greater age at the time symptoms in the joints first appeared; this age was, on average, ...

Mathon, G.; Pare?, C.; Me?nard, H.; Te?treault, L.; Camerlain, M.

1980-01-01

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HLA-B27 associated spondyloarthropathy, an autoimmune disease based on crossreactivity between bacteria and HLA-B27 ?  

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Most autoimmune diseases are associated with certain HLA types. Therefore, spondyloarthropathies (SpA) strongly associated with HLA-B27, are also often classified as autoimmune diseases. This study questions whether SpA indeed fulfils the criteria of an autoimmune disease. The Medline database was searched for all reports between 1966 and April 1998 on the presence of autoimmune reactivity in SpA patients. This search yielded 45 articles on this subject. Only eight articles study T cell re...

Ringrose, J. H.

1999-01-01

38

Polymorphonuclear cell motility, ankylosing spondylitis, and HLA B27.  

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Polymorphonuclear leucocyte (PMN) function was studied in 29 subjects with ankylosing spondylitis (AS). Of these, 20 were HLA B27+ve and 9 B27-ve. There were 30 controls and, of these, 15 were B27+ve. Random and directed cell migration was measured by 2 techniques: migration through a micropore filter and migration under an agar film. The chemo-attractant was either case in-activated serum or zymosan-activated serum. By both techniques directed motility was increased in subjects with B27 or w...

Pease, C. T.; Fordham, J. N.; Currey, H. L.

1984-01-01

39

HLA B27 allele types in homogeneous groups of juvenile idiopathic arthritis patients in Latvia  

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Abstract Juvenile idiopathic arthritis (JIA) is a heterogeneous condition and therapeutic strategies vary in different JIA types. The routinely accepted practice to start with Sulphasalazine (SS) as the first line treatment in patients with HLA B27 positive JIA proves to be ineffective in a large proportion of children. Objective to investigate HLA B27 positive JIA patients clinical characteristics, determined HLA B27 allele types and their connection with antirheumati...

Guseinova Dinara; Lazareva Arina; Sochnevs Arturs; Zavadska Dace; Eglite Jelena; Stanevicha Valda; Shantere Ruta; Gardovska Dace

2010-01-01

40

A subset of HLA-B27 molecules contains peptides much longer than nonamers.  

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An unusual monoclonal antibody (MARB4) directed against HLA-B27 that reacts with only approximately 5-20% of the cell surface HLA-B27 was used for large-scale purification of these molecules. Subsequent mass spectrometry of HLA-B27-bound peptides showed that the minor MARB4-reactive population contained peptides primarily from 900 to 4000 Da in size (approximately 8-33 amino acid residues), whereas the major HLA-B27 population contained peptides in the mass range of 900-1400...

Urban, R. G.; Chicz, R. M.; Lane, W. S.; Strominger, J. L.; Rehm, A.; Kenter, M. J.; Uytdehaag, F. G.; Ploegh, H.; Uchanska-ziegler, B.; Ziegler, A.

1994-01-01

 
 
 
 
41

Inhibiting HLA-B27 homodimer-driven immune cell inflammation in spondylarthritis.  

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OBJECTIVE: Spondylarthritides (SpA), including ankylosing spondylitis (AS), are common inflammatory rheumatic diseases that are strongly associated with positivity for the HLA class I allotype B27. HLA-B27 normally forms complexes with ?(2) -microglobulin (?(2) m) and peptide to form heterotrimers. However, an unusual characteristic of HLA-B27 is its ability to form ?(2) m-free heavy chain homodimers (HLA-B27(2) ), which, unlike classic HLA-B27, bind to killer cell immunoglobulin-like rece...

Payeli, Sk; Kollnberger, S.; Marroquin Belaunzaran, O.; Thiel, M.; Mchugh, K.; Giles, J.; Shaw, J.; Kleber, S.; Ridley, A.; Wong-baeza, I.; Keidel, S.; Kuroki, K.; Maenaka, K.; Wadle, A.; Renner, C.

2012-01-01

42

HLA B-27 Subtypes in Turkish Patients with Spondyloarthropathy and Healthy Controls  

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The frequency and the distribution of HLA-B27 subtypes in spondylarthropathy (SpA) patients and controls were investigated in a sample Turkish population. B27 subtyping was performed by PCR-SSP method in two groups: 49 unrelated HLA-B27 positive Turkish patients with the diagnosis of SpA according to the European Spondyloarthropathy Study Group Criteria, and 55 HLA-B27 positive healthy controls. The frequency of HLA-B?27 was 2.6% in the Turkish population, and B?2705 was the predominant a...

Fatma Savran Oguz; Lale Ocal; Ali Sarper Diler; Hilmi Ozkul; Faruk Asicioglu; Esen Kasapoglu; Gokay Bozkurt; Meral Konice; Mahmut Carin

2004-01-01

43

Cell-surface expression and immune receptor recognition of HLA-B27 homodimers.  

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OBJECTIVE: HLA-B27 is capable of forming in vitro a heavy-chain homodimer structure lacking beta(2)-microglobulin. We undertook this study to ascertain if patients with spondylarthritis express beta(2)-microglobulin-free HLA-B27 heavy chains in the form of homodimers and receptors for HLA-B27 homodimers. METHODS: Expression of HLA-B27 heavy chains by mononuclear cells was analyzed by fluorescence-activated cell sorter staining, Western blotting with the monoclonal antibody HC-10, and 2-dimens...

Kollnberger, S.; Bird, L.; Sun, My; Retiere, C.; Braud, Vm; Mcmichael, A.; Bowness, P.

2002-01-01

44

Klebsiella 'modifying factor': binding studies with HLA-B27+ and B27- lymphocytes.  

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On the basis that extracts of some klebsiella organisms bind selectively to the lymphocytes of HLA-B27+ individuals and induce the appearance of new antigens, attempts were made to detect the binding of klebsiella products to HLA-B27+ and B27- lymphocytes by a number of different techniques. Firstly, blocking of the binding of two different HLA-B27 specific monoclonal antibodies to HLA-B27+ lymphocytes has been examined following exposure of the lymphocytes to a cell-free culture filtrate fro...

Trapani, J. A.; Mckenzie, I. F.

1985-01-01

45

The arthritis-associated HLA-B*27:05 allele forms more cell surface B27 dimer and free heavy chain ligands for KIR3DL2 than HLA-B*27:09  

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OBJECTIVES: HLA-B*27:05 is associated with AS whereas HLA-B*27:09 is not associated. We hypothesized that different interactions with KIR immune receptors could contribute to the difference in disease association between HLA-B*27:05 and HLAB*27:09. Thus, the objective of this study was to compare the formation of ?2m-free heavy chain (FHC) including B27 dimers (B272) by HLA-B*27:05 and HLA-B*27:09 and their binding to KIR immunoreceptors. METHODS: We studied the formation of HLA-B*27:05 and ...

Cauli, A.; Shaw, J.; Giles, J.; Hatano, H.; Rysnik, O.; Payeli, S.; Mchugh, K.; Dessole, G.; Porru, G.; Desogus, E.; Fiedler, S.; Ho?lper, S.; Carette, A.; Blanco-gelaz, Ma; Vacca, A.

2013-01-01

46

Monocyte-derived dendritic cells from HLA-B27.  

Science.gov (United States)

IntroductionThis study aimed to compare the functional capacity and gene expression profile of monocyte-derived dendritic cells (MD-DCs) in HLA-B27+ axial spondyloarthritis (SpA) patients and healthy controls.MethodsMD-DCs were differentiated with interleukin 4 (IL-4) and granulocyte-macrophage colony-stimulating factor (GM-CSF) for seven days, starting from purified CD14+ monocytes and stimulated with lipopolysaccharide (LPS) for six and twenty four hours. Their capacity to stimulate allogeneic CD4+ T cells from unrelated healthy donor was tested. Transcriptomic study was performed with Affymetrix HuGene 1.0 ST microarrays. Gene expression levels were compared between patients and controls using a multivariate design under a linear model (LIMMA). Real-time quantitative PCR (qRT-PCR) was performed for validation of the most striking gene expression differences.ResultsThe stimulatory capacity of allogeneic CD4+ T cells by MD-DCs from SpA patients was decreased. Transcriptomic analysis revealed 81 genes differentially expressed in MD-DCs between SpA patients and controls (P 1.5). Four selected genes were validated by qRT-PCR: ADAMTS15, CITED2, F13A1 and SELL. Expression levels of ADAMTS15 and CITED2, encoding a metallopeptidase and a transcription factor, respectively, were inversely correlated with each other (R¿=¿0.75, P¿=¿0.0003). Furthermore, in silico analysis identified several genes of the Wnt signaling pathway having expression co-regulated with CITED2.ConclusionThis study revealed altered function and gene expression pattern in MD-DCs from HLA-B27+ axial SpA. Co-expression study showed an inverse correlation between ADAMTS15 and CITED2. Moreover, the Wnt signaling pathway appeared as deregulated in SpA MD-DCs, a finding which may be connected to Th17-driven inflammatory responses. PMID:25142923

Talpin, Alice; Costantino, Félicie; Bonilla, Nelly; Leboime, Ariane; Letourneur, Franck; Jacques, Sébastien; Dumont, Florent; Amraoui, Sonia; Dutertre, Charles-Antoine; Garchon, Henri-Jean; Breban, Maxime; Chiocchia, Gilles

2014-08-21

47

Oral treatment with recombinant human interleukin-11 improves mucosal transport in the colon of human leukocyte antigen-B27 transgenic rats.  

Science.gov (United States)

Recombinant human interleukin (IL)-11 is a pleiotropic cytokine with anti-inflammatory activity. The objective of the study was to investigate whether oral treatment with rhIL-11 improves colonic epithelial dysfunction in the human leukocyte antigen (HLA)-B27 transgenic rat model of spontaneous chronic inflammation. Experiments were performed using adult male HLAB27 rats, whereas healthy nontransgenic F344 rats served as controls. Enteric-coated rhIL-11 multi-particles (equivalent to 500 microg/kg rhIL11) or placebo (formulation lacking rhIL-11) were administrated orally on alternate days for 2 weeks to HLA-B27 or F344 rats. Stool character was observed daily during the treatment period. Animals were euthanized at the end of treatment and colonic inflammation was evaluated be measuring tissue myeloperoxidase (MPO) activity. Epithelial transport in isolated colonic mucosal sheets was studied in modified Ussing chambers. Oral treatment of HLA-B27 rats with rhIL-11 reduced MPO activity in the colon and suppressed the clinical signs of diarrhea. The electrophysiological characteristics of mucosal transport were improved in the HLA-B27 rats treated with rhIL-11 compared with placebo. After rhIL-11 treatment the basal transepithelial resistance and the estimated paracellular resistance were significantly increased, neurally mediated secretory responses to electrical field stimulation were improved, and cholinoceptor sensitivity was normalized. Treatment with rhIL-11 had no significant effect on basal short circuit current and the maximal secretory response to carbachol or substance P. Our data demonstrate that oral rhIL-11 therapy is associated with suppression of mucosal inflammation and a concomitant improvement of epithelial resistance and neurally mediated secretion in a model of chronic HLA-B27 colitis. PMID:14569059

Venkova, Kalina; Keith, James C; Greenwood-Van Meerveld, Beverley

2004-01-01

48

HLA-B27 frequency in a group of patients with psoriatic arthritis Freqüência de HLA-B27 em uma amostra de pacientes com artrite psoriática  

Directory of Open Access Journals (Sweden)

Full Text Available BACKGROUND: HLA-B27 is associated with spondyloarthritis, a group of diseases that includes psoriatic arthritis. OBJECTIVES: To describe the HLA-B27 frequency in a group of Brazilian patients with psoriatic arthritis and correlate its presence or absence with their clinical manifestations. METHODS: Cross-sectional study with 44 psoriatic arthritis patients of a Rheumatology clinic. Demographic and social data were recorded, as were skin and joints clinical examination. HLA-B27 was tested. All data were processed descriptively and comparatively by appropriate software. Parametric and non parametric tests were used with 5% statistical significance. RESULTS: HLA-B27 was negative in 32 of the 44 patients (72,7%. Most of them were male, Caucasian, living in Rio de Janeiro, with plaque type psoriasis and average age of 52,9 years. There was statistical significant correlation between positive HLA-B27 and male gender (p=0,004. Negative HLA-B27 had a tendency to correlate with hands and wrists arthritis (p=0,07. There was an inverse significant correlation between HLA values and Schöber's test (p=0,02. CONCLUSION: Although HLA-B27 is negative in most of patients, it is significantly associated to male gender and inversely correlated with Schöber's test.FUNDAMENTOS: O HLA-B27 está associado às espondiloartrites, grupo de doenças que engloba, entre outras, a artrite psoriásica. OBJETIVOS: Descrever a freqüência de HLA-B27 em uma amostra de pacientes brasileiros com artrite psoriásica e correlacionar sua presença ou ausência com as manifestações clínicas dos mesmos. MÉTODOS: Estudo transversal avaliando 44 pacientes com artrite psoriásica de um ambulatório de Reumatologia. A avaliação consistia em registro de informações demográficas e sociais, exame clínico da pele e das articulações e pesquisa de HLA-B27. Os dados gerados foram tratados por meio de estatística descritiva e comparativa em Software apropriado. Foram utilizados testes paramétricos e não-paramétricos com significância estatística de 5%. RESULTADOS: O HLA-B27 resultou negativo em 32 dos 44 pacientes estudados (72,7%. A maioria dos pacientes era do sexo masculino, da raça branca, procedente do Rio de Janeiro, portador de psoríase em placas e com idade média de 52,9 anos. Houve associação estatisticamente significativa entre o HLA-B27 positivo e o sexo masculino (p=0,004. O HLA-B27 negativo teve tendência à correlação com artrite de mãos e punhos (p=0,07. Houve correlação inversa significativa entre os valores do HLA e do teste de Schöber (p=0,02. CONCLUSÃO: Apesar do HLA-B27 ser negativo na maioria dos pacientes estudados, esteve significativamente associado ao sexo masculino e inversamente correlacionado ao teste de Schöber.

Danilo Garcia Ruiz

2012-12-01

49

Development of a PCR method to detect HLA-B27 in ankylosing spondylitis  

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The aim of the project was to develop a PCR method to detect HLA-B27 at the Immunology Department of St. James hospital in Dublin. The HLA-B27 gene is common among patients with ankylosing spondylitis (AS). Ninety percent of patients with AS have the HLA-B27 gene and it is therefore counted as a risk factor and could be used as part of the diagnosis. Twenty-two frozen blood samples from patients with AS or suspected AS were donated from the rheumatology department at St. James hospital. PCR i...

Na?tterkvist, Ylva

2012-01-01

50

Functional Interaction of the Ankylosing Spondylitis-associated Endoplasmic Reticulum Aminopeptidase 1 Polymorphism and HLA-B27 in Vivo*  

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The association of ERAP1 with ankylosing spondylitis (AS)1 among HLA-B27-positive individuals suggests that ERAP1 polymorphism may affect pathogenesis by altering peptide-dependent features of the HLA-B27 molecule. Comparisons of HLA-B*27:04-bound peptidomes from cells expressing different natural variants of ERAP1 revealed significant differences in the size, length, and amount of many ligands, as well as in HLA-B27 stability. Peptide analyses suggested that the mechanism of ERAP1/HLA-B27 in...

Garci?a-medel, Noel; Sanz-bravo, Alejandro; Nguyen, Dung; Galocha, Begon?a; Go?mez-molina, Patricia; Marti?n-esteban, Adria?n; Alvarez-navarro, Carlos; Castro, Jose? A. Lo?pez

2012-01-01

51

Clinical Features and Prognosis of HLA-B27 Positive and Negative Anterior Uveitis in a Korean Population  

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Clinical features and prognosis of HLA-B27 positive anterior uveitis (AU) were assessed compared with HLA-B27 negative AU in a Korean population, based on the medical records of AU patients seen at a university hospital. Twenty-seven HLA-B27 negative, idiopathic AU patients (group I) and 55 HLA-B27 positive AU patients (group II) were studied. HLA-B27 positive group was further divided into 29 with associated systemic disease (seronegative spondyloarthropathy) (group IIA) and 26 without assoc...

Park, Sung Chul; Ham, Don-il

2009-01-01

52

The solvent-inaccessible Cys67 residue of HLA-B27 contributes to T cell recognition of HLA-B27/peptide complexes.  

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Crystallographic studies have suggested that the cysteine at position 67 (Cys(67)) in the B pocket of the MHC molecule HLA-B*2705 is of importance for peptide binding, and biophysical studies have documented altered thermodynamic stability of the molecule when Cys(67) was mutated to serine (Ser(67)). In this study, we used HLA-B27.Cys(67) and HLA-B27.Ser(67) tetramers with defined T cell epitopes to determine the contribution of this polymorphic, solvent-inaccessible MHC residue to T cell rec...

Appel, H.; Kuon, W.; Kuhne, M.; Hu?lsmeyer, M.; Kollnberger, S.; Kuhlmann, S.; Weiss, E.; Zeitz, M.; Wucherpfennig, K.; Bowness, P.; Sieper, J.

2004-01-01

53

Complete heart block in HLA B27 associated disease. Electrophysiological and clinical characteristics.  

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A genetic predisposition associated with HLA B27 for developing complete heart block with or without clinical or radiological signs of associated rheumatic disease has recently been found. In this electrophysiological study of 12 patients with spontaneous complete heart block and HLA B27 associated disease, of whom eight had ankylosing spondylitis, 10 had suprahisian second or third degree atrioventricular block (eight spontaneously and two during atrial pacing at rates below 90 impulses per ...

Bergfeldt, L.; Vallin, H.; Edhag, O.

1984-01-01

54

Formation of HLA-B27 homodimers and their relationship to assembly kinetics  

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The human HLA-B27 class I molecule exhibits a strong association with the inflammatory arthritic disorder ankylosing spondylitis and other related arthropathies. Major histocompatibility complex class I heavy chains normally associate with beta(2)-microglobulin and peptide in the endoplasmic reticulum before transit to the cell surface. However, an unusual characteristic of HLA-B27 is its ability to form heavy chain homodimers through an unpaired cysteine at position 67 in the peptide groove....

Antoniou, A. N.; Ford, S.; Taurog, J. D.; Butcher, G. W.; Powis, S. J.

2004-01-01

55

Absence of a specific effect of free radicals on HLA-B27.  

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The spondylitis associated HLA-B27 epitope includes a characteristic unpaired cysteine at amino acid position 67. On some B27 molecules the thiol (-SH) side chain of this residue seems to be available for chemical interactions. The possibility that free radicals produced during inflammation might specifically affect this group was investigated in this work. Cells bearing HLA-B27 were exposed to free radicals generated by ultraviolet irradiation or hydrogen peroxide, and HLA antigens were then...

Maclean, I. L.; Lowdell, M. W.; Blake, D. R.; Lunec, J.; Archer, J. R.

1992-01-01

56

HLA B27 allele types in homogeneous groups of juvenile idiopathic arthritis patients in Latvia  

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Full Text Available Abstract Juvenile idiopathic arthritis (JIA is a heterogeneous condition and therapeutic strategies vary in different JIA types. The routinely accepted practice to start with Sulphasalazine (SS as the first line treatment in patients with HLA B27 positive JIA proves to be ineffective in a large proportion of children. Objective to investigate HLA B27 positive JIA patients clinical characteristics, determined HLA B27 allele types and their connection with antirheumatic treatment in homogenous patient groups. Materials and methods 56 patients diagnosed with JIA and observed over the period 2006 to 2009 included in the study. HLAB27 allele types were determined using PCR method. Results In HLA B27 positive JIA patients mean disease onset was 12.34 ± 3.3 years. Most common (44% JIA type was enthesitis related arthritis. Positive response to the treatment with SS was found in 32% of patients, Methotrexate (MTX - in 43%, combined treatment - SS with MTX was effective in 12.5%. 12.5% of patients required combination MTX with Enbrel. Eight HLA B27 allele types were found in JIA patients in Latvia: *2702, *2703, *2704, *2705, *2710, *2715, *2717, *2728. The most common was *2705 - in 55% of cases. Among all the patients enthesitis related arthritis most commonly occurred in patients with HLAB*2705 allele (OR = 2.01, p Conclusions There are 8 different HLA B27 alleles in JIA patients in Latvia and the most common is *2705, but in order to assert them to be disease associated alleles, more extensive studies are needed, including control group of HLA B27 positive healthy individuals. Standard treatment approach with SS proves to be unsatisfactory in the majority of JIA patients. To improve children's quality of life achieving rapid disease control, the first line treatment in HLA B27 positive patients should be MTX. In order to start with the most appropriate drug it is necessary to determine HLAB 27 type at the onset of disease.

Guseinova Dinara

2010-10-01

57

Finnish HLA studies confirm the increased risk conferred by HLA?B27 homozygosity in ankylosing spondylitis  

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OBJECTIVE: To determine the influence of HLA-B27 homozygosity and HLA-DRB1 alleles in the susceptibility to, and severity of, ankylosing spondylitis in a Finnish population. METHODS: 673 individuals from 261 families with ankylosing spondylitis were genotyped for HLA-DRB1 alleles and HLA-B27 heterozygosity/homozygosity. The frequencies of HLA-B27 homozygotes in probands from these families were compared with the expected number of HLA-B27 homozygotes in controls under Hardy-Weinberg equilibri...

Jaakkola, E.; Herzberg, I.; Laiho, K.; Barnardo, Mc; Pointon, Jj; Kauppi, M.; Kaarela, K.; Tuomilehto-wolf, E.; Tuomilehto, J.; Wordsworth, Bp; Brown, Ma

2005-01-01

58

Genética, HLA-B27 y espondilitis anquilosante: 40 años / Genetics of ankylosing spondylitis  

Scientific Electronic Library Online (English)

Full Text Available SciELO Chile | Language: Spanish Abstract in spanish [...] Abstract in english Ankylosing spondylitis (AS) is a prototypical inflammatory disease of the locomotor system affecting axial skeleton. It is part of the general group of spondyloarthopathies (SpA). Its strong association with histocompatibility antigen HLA-B27 is known since 1973. However, HLA-B27 contribution to AS [...] genetic risk is approximately 16%. Therefore, other genes are necessarily involved in the pathogenesis of the disease. Genomic development and the possibility of making genome wide screening have contributed enormously to the study of the disease. In this paper, we describe the actual knowledge about AS genetic risk, which has contributed to understand the influence of HLA-B27 on the etiology and pathogenesis of the disease. We also intend to foresee how these findings will result in an improvement of patients’ quality of life.

Patricia, Castro-Santos; Miguel A, Gutiérrez; Roberto, Díaz-Peña.

1165-11-01

59

Determination of HLA-B27 Subtypes in Iranian Patients with Ankylosing Spondylitis  

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Full Text Available The human leukocyte antigen-B27 is one of the class I molecules of the major histocompatibility complex which is strongly associated with ankylosing spondylitis (AS. The strength of the disease association with B27 varies markedly among racial and ethnic populations. It is an allele family, which constitutes about 31 subtypes, with a considerable geographic and ethnic difference in distribution. It is important to know whether certain subtypes show any preferential association with AS. Because there is no report regarding HLA-B27 subtypes in Iranian patients with AS, the factthe main there are rarelystudies (if any; purpose of the present study was to assess the frequency of subtypes of human leukocyte antigen (HLA-B27 in patients with ankylosing spondylitis in Iranian populationOne hundred and nineteen AS patients (82 HLA-B27 positive and 37 HLA-B27 negative were selected for this study. HLA-B27 positive patients were selected screened for B*27 subtyping were performed by polymerase chain reaction amplification with sequence-specific primers (PCR-SSP for B*27 subtyping.. The results of present study revealed that onlythat only two subtypes were detected in Iranian patients, including: B*2705 (52 patients, 63.4% and B*2702 (30 patients, 36.6%. Our results showed a restricted number of HLA-B27 subtypes associated with AS in Iran and an elevated frequency of the B*2705 allele in these patients similar to other Euro-Caucasoid (Aryan groups in the world.

Behrooz Nikbin

2008-05-01

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The biochemistry and immunology of non-canonical forms of HLA-B27  

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HLA-B27 (B27) is strongly associated with the spondyloarthritides. B27 is expressed at the cell surface of antigen presenting cells (APC) both as canonical ?2m-associated and non-canonical ?2m-free heavy chain (FHC) forms which include B27 dimers (termed B272). B27 FHC forms arise in an endosomal compartment from recycling ?2m-associated B27. Formation of cell surface FHC dimers is critically dependent on an unpaired reactive cysteine 67 in the ?1 helix of the class I heavy chain. HLA-B27...

Shaw, J.; Hatano, H.; Kollnberger, S.

2014-01-01

 
 
 
 
61

HLA class I typing by PCR: HLA-B27 and an African B27 subtype.  

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We describe a rapid method of HLA class I typing using the polymerase chain reaction and oligonucleotide hybridisation that eliminates the requirements for viable lymphocytes and allows subtypes to be defined. We have used this to demonstrate that the predominant subtype of HLA-B27 in the Gambia, West Africa, is HLA-B*2703, which is very rare or absent in other racial groups. This subtype differs from the common Caucasian HLA-B27 subtypes in its recognition by cytotoxic T cells. We propose th...

Hill, Av; Allsopp, Ce; Kwiatkowski, D.; Anstey, NM; Greenwood, BM; Mcmichael, Aj

1991-01-01

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HLA-B27 (antigen in retroperitoneal fibrosis in a family  

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Full Text Available Idiopathic retroperitoneal fibrosis is a rare disease of undetermined aetiology. It is important to distinguish this entity from retroperitoneal fibrosis secondary to malignancy or specific inflammatory disease. There have been no prior means of excluding this condition without surgical exploration and histopathologic study of the excised tissue. A genetic predisposition is suggested for the development of idiopathic primary retroperitoneal fibrosis in patients who are HLA-B27 antigen positive. In this study we present three cases of idiopathic retroperitoneal fibrosis in a family (2 brothers and their grandfather. The presence of HLA-B27 antigen positivity was identified in two of them.

Mohammad Yazdani

2007-03-01

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Use of HLA-B27 tetramers to identify low-frequency antigen-specific T cells in Chlamydia-triggered reactive arthritis.  

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Reports of the use of HLA-B27/peptide tetrameric complexes to study peptide-specific CD8+ T cells in HLA-B27+-related diseases are rare. To establish HLA-B27 tetramers we first compared the function of HLA-B27 tetramers with HLA-A2 tetramers by using viral epitopes. HLA-B27 and HLA-A2 tetramers loaded with immunodominant peptides from Epstein–Barr virus were generated with comparable yields and both molecules detected antigen-specific CD8+ T cells. The application of HLA-B27 tetramers in HL...

Appel, H.; Kuon, W.; Kuhne, M.; Wu, P.; Kuhlmann, S.; Kollnberger, S.; Thiel, A.; Bowness, P.; Sieper, J.

2004-01-01

64

Evaluation of 278 hla-b27 positive patients suspected of seronegative spondyloarthropathies  

International Nuclear Information System (INIS)

To determine HLA-B27 prevalence in patients suspected of Seronegative spondyloarthropathy referred to the Transplantation Department of Blood Transfusion Organization, and to evaluate clinical findings among HLA-B27 positive patients. One thousand six hundred ten patients having clinical manifestation of seronegative SpAs were screened for HLA typing by serological methods from January 1997 to June 2002 at Transplantation Department of Blood Transfusion Organization, Ahwaz, Iran. Serologic-based HLA typing using Antigen-specific sera to determine a person's HLA type was performed. Among these patients, individuals found HLA-B27 positive were investigated regarding clinical findings, age, and sex distribution. In this study the frequency of HLA-B27 antigen was 17.26% (278 cases). The minimum age in males was 10 years and the maximum age in female was 70 years. Median age with seronegative SpAs findings (34.2% including 28.42% females, 71.57% males) was 20-30 years. Based on our results, the most frequent clinical manifestation, was peripheral joints arthritis (58.7%; 34.35% females, 65.65 % males). There were no association between any of the major clinical manifestations and age or sex distribution. These findings confirm the strong association of the HLA B27 allele with various types of spondyloarthritis and suggests that HLA typing would help in the diagnosis of seronagative SpAs, specially ankylosing spondylitis with indeterminate clinical presentation and also in rminate clinical presentation and also in identifying at risk family members. (author)

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Interaction of HLA-B27 homodimers with KIR3DL1 and KIR3DL2, unlike HLA-B27 heterotrimers, is independent of the sequence of bound peptide.  

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HLA-B27 can form beta-2 microglobulin (beta2m)-associated heterotrimers (HLA-B27) and beta2m-free homodimers (B27(2)). Here, we study the role of complexed peptide in the interaction of these forms of B27 with the killer cell immunoglobulin (Ig)-like receptors KIR3DL1 and KIR3DL2 and with Ig-like transcripts LILRB1 and LILRB2. HLA-B27 tetramers complexed with three of five different naturally processed self peptides and three of seven pathogen-derived epitopes bound to KIR3DL1-expressing tran...

Kollnberger, S.; Chan, A.; Sun, My; Chen, Ly; Wright, C.; Di Gleria, K.; Mcmichael, A.; Bowness, P.

2007-01-01

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Autoantibodies to HLA B27 in the sera of HLA B27 patients with ankylosing spondylitis and Reiter's syndrome. Molecular mimicry with Klebsiella pneumoniae as potential mechanism of autoimmune disease  

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Ankylosing spondylitis (AS) and Reiter's syndrome (RS) both show a strong correlation with the HLA B27 haplotype. We studied whether sharing of homologous amino acid sequences in the HLA B27 antigen with an invading microbe might occur, and if so, what is the biological significance of such homology. In a computer search of the Dayhoff data bank, we found a homology of six consecutive amino acids between HLA B27.1 antigen residues 72-77 and Klebsiella pneumoniae nitrogenase residues 188-193. ...

1987-01-01

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IgG and IgA immune response against klebsiella in HLA-B27-associated anterior uveitis.  

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Enteric infections with Gram-negative bacteria are thought to play an important part in HLA-B27-associated disease such as Reiter's syndrome and reactive arthritis. But the role of bacterial infections in HLA-B27-positive ankylosing spondylitis (AS) and acute anterior uveitis (AU) is still controversial. A special interest has recently been devoted to the role of klebsiella infection in HLA-B27-associated disease. We studied the humoral immune response against a 'cross-reactive' strain of Kle...

Kijlstra, A.; Luyendijk, L.; Gaag, R.; Kregten, E.; Linssen, A.; Willers, J. M.

1986-01-01

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Cutting edge: HLA-B27 can form a novel beta 2-microglobulin-free heavy chain homodimer structure.  

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HLA-B27 has a striking association with inflammatory arthritis. We show that free HLA-B27 heavy chains can form a disulfide-bonded homodimer, dependent on residue Cys67 in their extracellular alpha 1 domain. Despite the absence of beta 2-microglobulin, HLA-B27 heavy chain homodimers (termed HC-B27) were stabilized by a known peptide epitope. HC-B27 complexes were recognized by the conformation-specific Ab W6/32, but not the ME1 Ab. Surface labeling and immunoprecipitation demonstrated the pre...

Allen, Rl; O Callaghan, Ca; Mcmichael, Aj; Bowness, P.

1999-01-01

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A homogeneous HLA-B*27 genotyping assay using dried reagent mixtures.  

Science.gov (United States)

The presence of HLA-B*27 allele with patients suspected with ankylosing spondylitis can be used in the diagnostic process. We have developed an assay for typing for the HLA-B*27 in whole blood dried on sample collection cards using pre-dried reagent wells and homogeneous time-resolved fluorescence based PCR approach. Essentially only the sample needs to be added to the dry ready-to-use reaction well in order to start the homogenous amplification assay. The method was validated with 229 samples also typed with an existing DELFIA-based method and results of both assays were 100% concordant. The dried reagents were shown to be stable at least up to eight weeks at room temperature without any decline in their performance. PMID:19893203

Kiviniemi, Minna; Ilonen, Jorma; Lövgren, Timo

2009-01-01

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Structural analysis of an HLA-B27 functional variant: identification of residues that contribute to the specificity of recognition by cytolytic T lymphocytes.  

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The structure of a variant HLA-B27 antigen, B27.2, that is distinguished from the HLA-B27.1 and HLA-B27.3 subgroups by specific cytolytic T lymphocytes has been established by comparative peptide mapping and sequence analysis. There are only three amino acid substitutions between B27.1 and B27.2: aspartate-77, threonine-80, and leucine-81 in HLA-B27.1 are changed to asparagine-77, isoleucine-80, and alanine-81 in HLA-B27.2. These changes account for their single charge difference detectable b...

Vega, M. A.; Ezquerra, A.; Rojo, S.; Aparicio, P.; Bragado, R.; Lo?pez Castro, J. A.

1985-01-01

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HLA-B27 and its Associated Clinical and Biochemical Presentation among Ghanaians with Ankylosing Spondylitis  

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Full Text Available HLA-B27 is a genetic predisposition marker for the development of Ankylosing Spondylitis (AS. AS is uncommon in West-Africa, but due to environmental and lifestyle changes, its prevalence is said to be increasing. This study sought to determine the baseline prevalence of HLA-B27 among Ghanaians presenting with AS, find out their disease activity, clinical presentation, presence of extra-articular manifestations, inflammation and dyslipidemia. In a cross-sectional study, 65 AS subjects were recruited from the orthopaedic departments of two leading Teaching Hospitals and a private laboratory, medilab diagnostics with centres across the country. Fifty healthy blood donors were also recruited as control group. HLA-B27, BASDAI score, Lipid profile, TNF-? and ESR levels were estimated among them. Statistical comparisons were analyzed using the one way ANOVA followed by Bonferroni’s Multiple Comparison test. There were four HLA-B27 positives representing 4.6%, the mean BASDAI score was 44.7/100. 48 AS patients had Sacroiliitis in their X-ray reports. None had a family history or any extra-articular manifestations. AS subjects had higher (p-1 compared to 5.70±0.48 pg mL-1 of control whiles the ESR was 34.64±1.87 mm h-1 as compared to 9.23±0.91 mm h-1 of controls. AS patients had moderate disease activity with no extra articular manifestation and a prevalence of 4.6%. Dyslipidemia was prominent and that inflammation plays a pivotal role in the development of atherosclerosis.

Robert E. Quansah

2012-01-01

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Intravitreal Triamcinolone Acetonide for Macular Edema in HLA-B27 Negative Ankylosing Spondylitis  

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We report a case of a human leukocyte antigen B27 (HLA-B27)-negative patient with cystoid macular edema (CME) and ankylosing spondylitis (AS) after treatment with triamcinolone acetonide. The patient complained of deterioration of visual acuity of the right eye during the last 10 days. At presentation visual acuity of the right eye was 0.2, and the ophthalmic examination did not reveal any sign of active uveitis. Fluorescein angiography (FA) and ocular coherent tomography (OCT) showed CME. Th...

Moschos, M. M.; Gatzioufas, Z.; Margetis, I.

2010-01-01

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Different HLA-B27 subtypes present the same immunodominant Epstein-Barr virus peptide  

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An immunological basis has been postulated for the strong association between at least five subtypes of the HLA-B27 allele (B27.01, .02, .04, .05, and .06) and ankylosing spondylitis, namely that cytotoxic T lymphocyte (CTL) responses are induced against an "arthritogenic" peptide that these different subtypes can all present. This requires a degree of overlap between the peptide binding repertoires of different B27 molecules. The present work, using CTL responses to Epstein-Barr virus (EBV) ...

1993-01-01

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Bilateral macular thickening in mild unilateral anterior uveitis: is HLA-B27 involved?  

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Abstract Background Macular thickening (MT) without clinically recognized macular edema has been described in anterior uveitis (AU). Although fellow-eyes of patients have been used as controls in several studies, little is known about macular thickness in these eyes. We studied the rate and extent of MT in both AU-affected and quiescent fellow-eyes of phakic AU patients with good visual acuity (VA). We also assessed macular thickness related to HLA-B27 presence and to recurre...

Wexler Alexandra; Sand Trond; Elsås Tor B

2012-01-01

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The role of HLA-B27 molecules in the pathogenesis of ankylosing spondylitis  

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Full Text Available Ankylosing Spondylitis (AS is characterised by the strongest association with an HLA antigen ever described for any disease. It represents therefore the ideal model for the understanding of the link between immune-mediated diseases and the HLA system. The role of HLA-B27 in the pathogenesis of AS will be discussed focusing on the recently described higher expression of these molecules in patients with AS compared with healthy controls.

R. Pala

2011-09-01

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2 cases of HLA-B27-positive seronegative spondylarthritides in pediatric age treated with adalimumab.  

Science.gov (United States)

Seronegative spondyloarthritis is strongly correlated to HLA-B27, and in the long term, it causes limitations to the movements of vertebral joints. In recent years, the numbers of patients diagnosed with axial spondyloarthritis have increased due to the widespread use of magnetic resonance imaging (MRI) for diagnostic imaging. We report the cases of 2 pediatric patients diagnosed with axial spondyloarthritis, and whose disease activity was successfully controlled using adalimumab. In case 1, the patient was a 15-year-old boy. The onset of the disease was marked by neck pain ; HLA-B27 was positive, and the MRI revealed sacroiliac arthritis. After being diagnosed with axial spondyloarthritis, he began receiving oral steroid therapy. Gradual recurrence was observed, and adalimumab treatment was initiated. In case 2, the patient was a 9-year-old boy. Bilateral pain was present in the shoulder joints, ankles, and knee joints. The patient was diagnosed with polyarticular juvenile idiopathic arthritis, and treatment using oral steroids, immunosuppressants and tocilizumab. The arthralgia disappeared, but at the age of 12 years, pain recurred in the sacroiliac joint and the Achilles tendon, the HLA-B27 was positive, and the MRI revealed sacroiliac arthritis. The condition was diagnosed as axial spondyloarthritis; adalimumab treatment was initiated. Adalimumab was effective in the treatment of axial spondylitis occurring in childhood. PMID:24390108

Sato, Tomomi; Nozawa, Tomo; Kanetaka, Taichi; Kikuchi, Masako; Sakurai, Nodoka; Yamazaki, Kazuko; Momoi, Takahiro; Namai, Yoshiyuki; Yokota, Shumpei

2013-01-01

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HLA-B27 predicts a more extended disease with increasing age at onset in boys with juvenile idiopathic arthritis  

DEFF Research Database (Denmark)

OBJECTIVE: Juvenile idiopathic arthritis (JIA) is a heterogeneous condition with very few clinical and laboratory signs that can help predict the course and severity of the disease in the individual patient. The cell-surface antigen HLA-B27 is well known to be associated with spondyloarthropathies, reactive arthritis, and enthesitis. HLA-B27 plays an important role in the classification of JIA, since evidence of sacroiliitis most often evolves after years of arthritis in other joints. We investigated the associations of HLA-B27 and the clinical manifestations of JIA using a method as close to a population-based study as possible. METHODS: We studied an incidence-based cohort of 305 patients collected prospectively in 3 Nordic countries (Sweden, Norway, Denmark). Clinical and serological data of the first 3 years of the disease were collected. RESULTS: HLA-B27 was found to be positive in 25.5% of the patients, and we found a higher proportion of HLA-B27-positive boys with older age at disease onset (p=0.034). Regression analysis showed a correlation of 0.7 in the HLA-B27-positive boys, pointing to a higher risk of more joint involvement with older age at disease onset. By Fisher's exact test, involvement of small joints in the lower extremities was associated with HLA-B27 in boys (p=0.011), but not in girls (p=0.687). HLA-B27 was associated with inflammatory back pain in both sexes (p=0.041 in boys, p=0.042 in girls), but with enthesitis only in boys (p<0.001 in boys, p=0.708 in girls). CONCLUSION: HLA-B27 is of increasing importance with older age at disease onset in boys with JIA, predicting more active joints within the first 3 years of disease, and also involving small joints in the lower extremity to a greater degree than in HLA-B27-negative boys. During the first 3 years of disease the occurrence of HLA-B27 is associated with inflammatory back pain in both sexes, but with enthesitis only in boys. Our data present new challenges for the ILAR classification of JIA Udgivelsesdato: 2008/10

Berntson, Lillemor; Damgård, Michael

2008-01-01

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Bilateral macular thickening in mild unilateral anterior uveitis: is HLA-B27 involved?  

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Full Text Available Abstract Background Macular thickening (MT without clinically recognized macular edema has been described in anterior uveitis (AU. Although fellow-eyes of patients have been used as controls in several studies, little is known about macular thickness in these eyes. We studied the rate and extent of MT in both AU-affected and quiescent fellow-eyes of phakic AU patients with good visual acuity (VA. We also assessed macular thickness related to HLA-B27 presence and to recurrence, since these issues have been almost unexplored by previous optical coherence tomography (OCT studies. Methods Patients with AU were prospectively included and macular thickness was measured with OCT initially and on follow up. Macular thickness in patients’ affected eyes (n?=?30 as well as in their quiet fellow-eyes (n?=?28 was compared with eyes of age- and gender matched controls. Inter-ocular differences in macular thickness between AU affected eyes and their fellow-eyes were assessed in patients (n?=?28, also in a subgroup with visual acuity???0.8 (n?=?23 by one-sample Student’s?t-tests. Inter-ocular differences were also assessed related to HLA-B27 presence and related to the status of current AU episode (initial or relapse. Results Subclinical MT is present in both quiet fellow-eyes and AU-affected eyes of patients. MT was found in most cases of AU, even in phakic eyes with good VA. There was a larger increase in macular thickness in HLA-B27-positive than in HLA-B27-negative patients. No differences in macular thickness were found between patients with their first AU episode and patients with recurrent episodes. Conclusions MT probably reflects systemic immune-mediated response to the inflammatory disorder in AU, and it is possible that HLA-B27-related factors are involved in the pathogenesis of AU. These observations are in line with and extend the current understanding of the mechanisms behind MT in AU.

Wexler Alexandra

2012-07-01

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Revisiting the arthritogenic peptide theory: Quantitative not qualitative changes in the peptide repertoire of HLA-B27 allotypes.  

Science.gov (United States)

Objective: The association of HLA-B27 with spondyloarthropathy is one of the strongest documented for any autoimmune disease. A common hypothesis for this association is the arthritogenic peptide concept. This dictates that differences in the peptide binding preferences of disease associated and non-associated HLA-B27 allotypes underpin the presentation of bacterial and self-peptides leading to cross-reactive T cell immunity and subsequent autoimmune attack of affected tissues. Methods: Qualitative differences in the peptides bound to the 8 most frequent HLA-B27 subtypes were determined by tandem mass spectrometry and quantitative changes in allelic binding specificities determined by highly sensitive and targeted multiple reaction monitoring mass spectrometry. Results: Here we identify over 7500 MHC class I peptides derived from the 8 most common HLA-B27 allotypes, HLA-B*27:02 to HLA-B*27:09. We describe individual binding motifs for these alleles for 9-12 mer ligands. The peptide repertoires of these closely related alleles show significant overlap. Allelic polymorphism resulting in changes in the amino acid composition of the antigen binding cleft manifests largely as quantitative changes in the peptide cargo of these molecules. Conclusion: Absolute binding preferences do not explain disease association. The arthritogenic peptide theory needs to be reassessed in terms of quantitative changes in self-peptide presentation, T cell selection and altered conformation of bound peptides. This article is protected by copyright. All rights reserved. PMID:25418920

Schittenhelm, Ralf B; Lim Kam Sian, Terry C C; Wilmann, Pascal G; Dudek, Nadine L; Purcell, Anthony W

2014-11-21

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A comparison of self-reported joint symptoms following infection with different enteric pathogens: effect of HLA-B27  

DEFF Research Database (Denmark)

OBJECTIVE: We conducted a case-case comparison study to estimate the attack-rate of reactive joint pain (JPrea) following intestinal infections, and evaluated whether the susceptibility and severity of joint symptoms was associated with the tissue-type HLA-B27. METHODS: Consecutive patients with positive fecal culture for Salmonella, Campylobacter, Yersinia, Shigella, and E. coli were addressed by questionnaires inquiring about gastrointestinal (GI) symptoms and the occurrence of joint pain in a previously healthy joint within 4 weeks after onset of infection. A blood sample was requested for HLA-B27 typing. RESULTS: Of 3146 patients invited, 2105 (67%) responded to the survey questionnaire. The triggering infections were Campylobacter, 1003; Salmonella, 619; E. coli, 290; Shigella, 102; and Yersinia, 91. JPrea was reported by 294 subjects: Campylobacter, 131 (13.1%); Salmonella, 104 (16.8%); Yersinia, 21 (23.1%); Shigella, 10 (9.8%); and E. coli, 28 (9.7%). There was a significant association between severity of gastroenteritis and development of arthralgia (p = 0.001). The odds ratio (OR) for JPrea in an HLA-B27-positive individual was 2.62 (95% CI 1.67-3.93) for the entire group. A significant association between JPrea and HLA-B27 was found for Salmonella, Shigella, and Yersinia; not, however, for Campylobacter and E. coli. HLA-B27-positive patients had a significantly increased risk for severe joint symptoms. CONCLUSION: Our study shows that JPrea after GI infection is positively correlated to severity of GI symptoms. HLA-B27 is not associated with joint pain after Campylobacter. Intestinal E. coli seems to be an arthritogenic pathogen. A significant association between HLA-B27 and severity of joint pain was observed Udgivelsesdato: 2008/3

Schiellerup, P.; Krogfelt, K.A.

2008-01-01

 
 
 
 
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HLA-B27 associated reactive spondyloarthropathies in a Dutch military hospital.  

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Forty-two male patients with reactive spondyloarthropathies in a Dutch military hospital are described with mean age of onset 21.9 years. Peripheral arthritis or sacroiliitis was present in all, eye symptoms in 21 (50%), and genitourinary disease in 15 (35.7%). Evidence of antecedent sexually acquired or enterocolitic infection was found only in three (7.2%). HLA-B27 antigen was detected in 34 (81%) of 42 patients. Additional data suggest that reactive spondyloarthropathies are the most commo...

Lionarons, R. J.; Zoeren, M.; Verhagen, J. N.; Lammers, H. A.

1986-01-01

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Observer variation in grading sacroiliac radiographs in HLA-B27 positive individuals  

International Nuclear Information System (INIS)

This study attempts to reconcile the apparent differences in the reported frequency of ankylosing spondylitis and radiological sacroilitis in HLA-B27 positive individuals. Pelvic radiographs from 125 Busselton subjects were mixed with 81 other films selected to illustrate the possible range of sacroiliac changes and were graded by observers who were involved in 2 of the conflicting studies and by a 3rd independent observer. Concordance was high for advanced bilateral disease but not for unilateral and milder changes. Variation between observers and the interpretation of sacroiliac radiographs is sufficiently large to account for much of the disagreement between frequency estimates

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Detection of HLA-B27 alleles by group-specific amplification and time-resolved fluorometry.  

Science.gov (United States)

This newly developed HLA-B27 assay combines a polymerase chain reaction (PCR) from blood spot samples with solution hybridisation in microtitration plate and with time-resolved fluorometry (TRF) as the detection system. In a multiplex amplification reaction, the 144 base pair region of HLA-B27 alleles is amplified with allele-specific primers simultaneously with the region of beta-actin gene as an internal control. Amplified products are collected onto streptavidin (SA)-coated microtitration wells, denatured and hybridised with a europium (Eu)-labelled HLA-B27 specific probe and a samarium (Sm)-labelled beta-actin specific probe. Finally, Eu and Sm fluorescence is enhanced and detected in a time-resolved fluorometer. The typing results obtained with 110 blood spot samples showed an exact match with serological class I HLA-typing. When this technique was further evaluated, 348 blood spot samples were clearly categorised into two populations, HLA-B27 positives and negatives. This new PCR-TRF method permits the automation of HLA-B27 assays and saves time and labour in routine diagnostics. PMID:9831394

Välimaa, L; Sjöroos, M; Luhtala, M; Toivanen, P; Lövgren, T; Ilonen, J

1998-10-01

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Endogenous Processing and Presentation of T-cell Epitopes from Chlamydia trachomatis with Relevance in HLA-B27-associated Reactive Arthritis*  

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Chlamydia trachomatis triggers reactive arthritis, a spondyloarthropathy linked to the human major histocompatibility complex molecule HLA-B27, through an unknown mechanism that might involve molecular mimicry between chlamydial and self-derived HLA-B27 ligands. Chlamydia-specific CD8+ T-cells are found in reactive arthritis patients, but the immunogenic epitopes are unknown. A previous screening of the chlamydial genome for putative HLA-B27 ligands predicted multiple peptides that were recog...

Cragnolini, Juan J.; Garci?a-medel, Noel; Lo?pez Castro, Jose? A.

2009-01-01

85

Two-dimensional gel analysis demonstrates no structural alteration of HLA-B27 polypeptides between patients with ankylosing spondylitis and healthy individuals.  

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After precipitation of the HLA-B27 antigen from the surface of peripheral blood lymphocytes (PBL) by means of an anti-HLA-B27 allospecific monoclonal antibody 2-dimensional gel electrophoresis was used to compare the structure of the B27 antigens derived from 5 patients with ankylosing spondylitis with that of healthy HLA-B27 positive counterparts. No significant difference in polypeptide structure was noted, which suggests that the pathogenesis of ankylosing spondylitis does not involve a st...

Trapani, J. A.; Walker, I. D.; Mckenzie, I. F.

1984-01-01

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Critical role of endoplasmic reticulum aminopeptidase 1 in determining the length and sequence of peptides bound and presented by HLA-B27.  

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OBJECTIVE: HLA-B27 and endoplasmic reticulum aminopeptidase 1 (ERAP1) are the two strongest genetic factors predisposing to ankylosing spondylitis (AS). A key aminopeptidase in class I major histocompatibility complex presentation, ERAP1 potentially contributes to the pathogenesis of AS by altering HLA-B27 peptide presentation. The aim of this study was to analyze the effects of ERAP1 on the HLA-B27 peptide repertoire and peptide presentation to cytotoxic T lymphocytes (CTLs). METHODS: ERAP1-...

Chen, L.; Fischer, R.; Peng, Y.; Reeves, E.; Mchugh, K.; Ternette, N.; Hanke, T.; Dong, T.; Elliott, T.; Shastri, N.; Kollnberger, S.; James, E.; Kessler, B.; Bowness, P.

2014-01-01

87

Multiple sclerosis and HLA-B27 negative sacroiliitis in a Crohn?s disease patient  

Directory of Open Access Journals (Sweden)

Full Text Available SUMMARY A relationship between inflammatory bowel disease and MS is supported by a higher than expected coexistence of these diseases among families and individuals. A 32 year-old male with Crohn?s disease of the terminal ileum diagnosed 4 years earlier and HLA-B27 bilateral sacroiliitis diagnosed two years earlier, was admitted to our hospital because of an acute episode of blurred vision. In addition the patient complained of urine incontinence. Before this admission the patient had been elsewhere administered three doses of Remicade and 16mg of methylprednisolone p.os. During admission the diagnosis of multiple sclerosis was made (MRI and IgG Index and Remicade was discontinued. The patient was started on therapy with interferon-beta for MS, oxybutynin hydrochloride (10mg/day for urine incontinence, prednizolone (10mg/day, methotrexate (10mg/week and azathioprine (100mg/day for Crohn?s disease and is now in excellent clinical status. To the best of our knowledge this is one of the very rare cases of Crohn?s disease with HLA-B27 negative sacroiliitis preceding multiple sclerosis diagnosis. Key words: Crohn?s disease, inflammatory bowel disease, ulcerative colitis, multiple sclerosis, Remicade

K.H. Katsanos, N. Tzambouras, E.V. Tsianos

2007-03-01

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Usage of Conventional PCR Technology for the Detection of HLA-B27 Allele: A Significant Molecular Marker of Ankylosing Spondylitis  

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Ankylosing spondylitis is a chronic inflammatory disease that has been linked to the human leukocyte antigen class I allele HLA-B27. More than 90 % of patients with ankylosing spondylitis possess the HLA-B27 allele, but only 1 % of people with HLA-B27 develop the disease. Ankylosing spondylitis predominately affects young males. The present study was planned to find out the involvement of HLA-B27 specific allele in relation to age and sex in symptomatic suspected patients of ankylosing spon...

Sharma, Narotam; Sharma, Veena; Masood, Tariq; Nautiyal, Satish Chandra; Sailwal, Shivani; Singh, Rajesh K.; Kushwaha, Rajeev K.; Singh, R. K.

2012-01-01

89

HLA-A*01:03, HLA-A*24:02, HLA-B*08:01, HLA-B*27:05, HLA-B*35:01, HLA-B*44:02, and HLA-C*07:01 Monochain Transgenic/H-2 Class I Null Mice : Novel Versatile Preclinical Models of Human T Cell Responses  

DEFF Research Database (Denmark)

We have generated a panel of transgenic mice expressing HLA-A*01:03, -A*24:02, -B*08:01, -B*27:05, -B*35:01, -B*44:02, or -C*07:01 as chimeric monochain molecules (i.e., appropriate HLA ?1?2 H chain domains fused with a mouse ?3 domain and covalently linked to human ?2-microglobulin). Whereas surface expression of several transgenes was markedly reduced in recipient mice that coexpressed endogenous H-2 class I molecules, substantial surface expression of all human transgenes was observed in mice lacking H-2 class I molecules. In these HLA monochain transgenic/H-2 class I null mice, we observed a quantitative and qualitative restoration of the peripheral CD8(+) T cell repertoire, which exhibited a TCR diversity comparable with C57BL/6 WT mice. Potent epitope-specific, HLA-restricted, IFN-?-producing CD8(+) T cell responses were generated against known reference T cell epitopes after either peptide or DNA immunization. HLA-wise, these new transgenic strains encompass a large proportion of individuals from all major human races and ethnicities. In combination with the previously created HLA-A*02:01 and -B*07:02 transgenic mice, the novel HLA transgenic mice described in this report should be a versatile preclinical animal model that will speed up the identification and optimization of HLA-restricted CD8(+) T cell epitopes of potential interest in various autoimmune human diseases and in preclinical evaluation of T cell-based vaccines.

Boucherma, Rachid; Kridane-Miledi, Hédia

2013-01-01

90

Dominant influence of an HLA-B27 restricted CD8+ T cell response in mediating HCV clearance and evolution.  

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Virus-specific CD8+ T cell responses play an important role in the natural course of infection; however, the impact of certain CD8+ T cell responses in determining clinical outcome has not been fully defined. A well-defined cohort of women inoculated with HCV from a single source showed that HLA-B27 has a strong association with spontaneous clearance. The immunological basis for this association is unknown. However, the finding is especially significant because HLA-B27 has also been shown to ...

Neumann-haefelin, C.; Mckiernan, S.; Ward, S.; Viazov, S.; Spangenberg, Hc; Killinger, T.; Baumert, Tf; Nazarova, N.; Sheridan, I.; Pybus, O.; Von Weizsa?cker, F.; Roggendorf, M.; Kelleher, D.; Klenerman, P.; Blum, He

2006-01-01

91

HLA-B27 associated cross-reactive marker on the cells of New Zealand patients with ankylosing spondylitis.  

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We have previously shown that antibodies raised in rabbits to certain enteric bacteria will specifically lyse, in a 51Cr release assay, the peripheral blood lymphocytes (PBL) of 80% of HLA-B27 positive patients with ankylosing spondylitis (B27+ AS+) but not the PBL of HLA-B27 positive normal controls (B27+ AS-). Other laboratories have been unable to reproduce these findings. This study was designed to ascertain whether this lack of reproducibility was due to a peculiarity of our B27+ AS+ pat...

Mcguigan, L. E.; Geczy, A. F.; Prendergast, J. K.; Edmonds, J. P.; Hart, H. H.; Bashir, H. V.

1986-01-01

92

Increased expression of human leucocyte antigen class I free heavy chains on monocytes of patients with spondyloarthritis and cells transfected with HLA-B27.  

Science.gov (United States)

Human leucocyte antigen (HLA)-B27 expression is correlated with spondyloarthritis (SpA), but its role in disease pathogenesis remains unclear. The aim of the study was to determine whether HLA-B27 free heavy chain (FHC) contributes to SpA pathogenesis. Flow cytometry was used to analyse the FHC expression on CD3+ and CD14+ cells in the peripheral blood (PB) and synovial fluid (SF) from SpA patients, healthy controls, and rheumatoid arthritis (RA) patients. Human monocytic U937 cell lines stably expressing enhanced green fluorescence protein (EGFP)/HLA-B27, EGFP/HLA-A2 or EGFP alone were created to further investigate the relation between HLA-B27 and FHC expression. The relative FHC level on CD14+ PB cells was significantly higher in SpA patients than in controls, but lower than on the SF cells of SpA patients. No significant correlation was found for relative FHC expression with HLA-B27 or ?2-microglobulin expression. HLA-B27-transfected U937 cells expressed higher FHC levels than either EGFP/HLA-A2- or EGFP-transfected cells. HLA class I FHC expression was significantly increased on monocytes of SpA patients and HLA-B27-transfected cells, implying that FHC, perhaps mostly derived from HLA-B27, plays an important role in SpA pathogenesis. PMID:25545293

Ding, Jin; Feng, Yuan; Zheng, Zhao Hui; Li, Xue Yi; Wu, Zhen Biao; Zhu, Ping

2015-02-01

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Comparación de la eficacia de la prednisolona y la rimexolona en el tratamiento de iridociclitis aguda en pacientes HLA-B27 positivos / Efficacy of prednisolone and rimexolone in HLA-B27 positive patients with acute anterior uveitis  

Scientific Electronic Library Online (English)

Full Text Available SciELO Mexico | Language: Spanish Abstract in spanish Objetivo: Comparar la eficacia y seguridad de las suspensiones oftálmicas de acetato de prednisolona al 1% y de rimexolona al 1%, en el tratamiento de la uveítis anterior aguda (UAA) en pacientes HLA B27+. Material y métodos: Se seleccionaron al azar 68 pacientes con UAA HLA -B27+ para tratamiento c [...] on acetato de prednisolona al 1% o rimexolona al 1%. En todos los pacientes la inflamación en cámara anterior era leve a moderada. La presión intraocular (PIO) y el grado de inflamación fueron evaluados semanalmentre durante seis semanas. Fue un estudio clínico prospectivo, aleatorio y doble ciego. Resultados: Al cuantificar las células en cámara anterior no se encontraron diferencias estadísticamente significativas entre uno y otro grupos. En el grupo de rimexolona el flare disminuyó desde la primera semana. En los grupos la presión intraocular se elevó con respecto a la basal desde la primera semana, la variación fue más significativa en el grupo de rimexolona. La PIO final fue menor en el grupo de rimexolona, siendo esta diferencia estadísticamente significativa . Conclusión: Para el tratamiento de la UAA HLA -B27+, leve a moderada, la rimexolona al 1% y la prednisolona al 1% tiene una eficiencia similar. En este estudio las variaciones de presión intraocular en los dos grupos no fueron clínicamente significativas. Abstract in english Purpose: To compare the efficacy and safety of prednisolone acetate 1 % vs. rimexolone 1 % ophthalmic suspension in the treatment of acute anterior uveitis (AAU) in HLA-B27+ patients. Methods: Sixty-eight AAU HLA-B27+ patients were randomly selected for treatment with prednisolone acetate 1% or Rime [...] xolone 1%. All patients showed mild to moderate anterior chamber inflammation. This was a prospective, randomized, double blind, clinical trial. Results: There was no statistically significant difference between both groups when anterior chamber cells were measured. In the rimexolone group, flare diminished since the first week. In both groups the intraocular pressure (IOP) raised since the first week; the increase washighly significant in the rimexolone group. Final intraocular pressure was higher in the prednisolone group. Conclusion: Rimexolone 1 % is as effective as prednisolone acetate 1% in the treatment of mild to moderate AAU HLA B27+. IOP increased in both groups, but this variation was not clinically significant.

Lourdes, Arellanes-García; Gonzalo, Padilla-Aguilar; Patricia, Navarro-López; Cynthia, Espinoza-Martínez.

2005-10-01

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Effects of tumor necrosis factor inhibitor on serum level of HLA-B27 and PDCD-1 in patients with ankylosing spondylitis.  

Science.gov (United States)

The aim of this study was to evaluate the effect of tumor necrosis factor-alpha (TNF-?) inhibitor-infliximab on ankylosing spondylitis (AS) patients and detect the serum level of HLA-B27 and PDCD-1 before and after TNF inhibitor treatment. 138 patients at active stage of AS were treated with infliximab; serum was collected before and after TNF-? inhibitor treatment for analysis. Reverse transcription-polymerase chain reaction (RT-PCR), flow cytometry, and enzyme-linked immuno sorbent assay were applied to detect the levels of HLA-B27 and PDCD-1 at different time points, which were used for statistical analysis with clinical data including two AS indicators (erythrocyte sedimentation rate--ESR and C-reactive protein--CRP). After the treatment for 6 weeks, RT-PCR showed that the gene expressions of HLA-B27 and PDCD-1 were significantly downregulated compared with baseline before infliximab treatment (P HLA-B27 and PDCD-1 double-labeled cells were significantly downregulated (P HLA-B27, and PDCD-1 of the AS patients were all significantly lower than the baseline levels (P HLA-B27 and PDCD-1 levels were all significantly correlated with ESR (P HLA-B27 and PDCD-1 in patients with AS. HLA-B27 and PDCD-1 are involved in the pathogenesis, and disease activities of AS. HLA-B27 and PDCD-1 are potentially the useful markers of AS activity and useful parameters to evaluate the effectiveness of anti-TNF-? inhibitor in treating AS. PMID:25005770

Chen, Xiaogang; Zhou, Xiaoqing; Li, Xia; Tang, Jinshan; Hu, Xiaowu; Wang, Junsheng; Xu, Cheng

2014-11-01

95

Autoantibodies to the HLA-B27 sequence cross-react with the hypothetical peptide from the arthritis-associated Shigella plasmid.  

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We previously reported elevated serum antibody levels to a peptide representing the HLA-B27 polymorphic region (B27 peptide) in HLA-B27(+) ankylosing spondylitis (AS) patients. A plasmid (pHS-2) isolated from arthritogenic Shigella flexneri strains had been shown to encode an amino acid sequence homologous to HLA-B27. Rabbit antibody to this sequence (pHS-2 peptide) strongly cross-reacted with B27 peptide and, to a much lesser extent, with Klebsiella nitrogenase peptide. Serum antibody levels...

Tsuchiya, N.; Husby, G.; Williams, R. C.; Stieglitz, H.; Lipsky, P. E.; Inman, R. D.

1990-01-01

96

In vitro mutagenesis of HLA-B27. Substitution of an unpaired cysteine residue in the alpha 1 domain causes loss of antibody-defined epitopes.  

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The HLA class I molecules identified serologically as HLA-B27 are highly associated with ankylosing spondylitis and related human disorders. All known HLA-B27 amino acid sequences contain a cysteine residue at position 67; no other published HLA class I sequence contains a cysteine within the hypervariable region of the alpha 1 domain, which extends from amino acid residues 63-84. To investigate the role of this cysteine residue in the antigenic structure of HLA-B27, we isolated a genomic clo...

Taurog, J. D.; El-zaatari, F. A.

1988-01-01

97

Antibody activity in ankylosing spondylitis sera to two sites on HLA B27.1 at the MHC groove region (within sequence 65-85), and to a Klebsiella pneumoniae nitrogenase reductase peptide (within sequence 181-199)  

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74 overlapping peptides of varying lengths from Klebsiella pneumoniae nitrogenase reductase (residues 181-199) and from the HLA B27.1 molecule (residues 65-85) were synthesized and tested by ELISA against sera from HLA B27+ ankylosing spondylitis (AS) patients, and sera from HLA B27+ and HLA B27- healthy first-degree relatives. Antibody activity in AS sera to Klebsiella peptides of four to eight amino acids was maximal with the peptide NSRQTDR. Activity to HLA B27 peptides was maximal with th...

1990-01-01

98

Structural analysis of an HLA-B27 functional variant, B27d detected in American blacks  

International Nuclear Information System (INIS)

The structure of a new functional variant B27d has been established by comparative peptide mapping and radiochemical sequencing. This analysis complete the structural characterization of the six know histocompatibility leukocyte antigen (HLA)-B27 subtypes. The only detected amino acid change between the main HLA-B27.1 subtype and B27d is that of Try59 to His59. Position 59 has not been previously found to vary among class I HLA or H-2 antigens. Such substitution accounts for the reported isoelectric focusing pattern of this variant. HLA-B27d is the only B27 variant found to differ from other subtypes by a single amino acid replacement. The nature of the change is compatible with its origin by a point mutation from HLB-B27.1. Because B27d was found only American blacks and in no other ethnic groups, it is suggested that this variant originated as a result of a mutation of the B27.1 gene that occurred within the black population. Structural analysis of B27d was done by comparative mapping. Radiochemical sequencing was carried out with 14C-labeled and 3H-labeled amino acids

99

Structural analysis of an HLA-B27 functional variant, B27d detected in American blacks  

Energy Technology Data Exchange (ETDEWEB)

The structure of a new functional variant B27d has been established by comparative peptide mapping and radiochemical sequencing. This analysis complete the structural characterization of the six know histocompatibility leukocyte antigen (HLA)-B27 subtypes. The only detected amino acid change between the main HLA-B27.1 subtype and B27d is that of Try/sub 59/ to His/sub 59/. Position 59 has not been previously found to vary among class I HLA or H-2 antigens. Such substitution accounts for the reported isoelectric focusing pattern of this variant. HLA-B27d is the only B27 variant found to differ from other subtypes by a single amino acid replacement. The nature of the change is compatible with its origin by a point mutation from HLB-B27.1. Because B27d was found only American blacks and in no other ethnic groups, it is suggested that this variant originated as a result of a mutation of the B27.1 gene that occurred within the black population. Structural analysis of B27d was done by comparative mapping. Radiochemical sequencing was carried out with /sup 14/C-labeled and /sup 3/H-labeled amino acids.

Rojo, S.; Aparicio, P.; Hansen, J.A.; Choo, S.Y.; Lopez de Castro, J.A.

1987-11-15

100

Microarray Analysis of Response of Salmonella during Infection of HLA-B27- Transfected Human Macrophage-Like U937 Cells  

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Full Text Available Abstract Background Human leukocyte antigen (HLA-B27 is strongly associated with the development of reactive arthritis (ReA in humans after salmonellosis. Human monocytic U937 cells transfected with HLA-B27 are less able to eliminate intracellular Salmonella enterica serovar Enteritidis than those transfected with control HLA antigens (e.g. HLA-A2. To investigate further the mechanisms by which HLA-B27-transfected cells allow increased replication of these bacteria, a DNA-based microarray was used for comparative genomic analysis of S. Enteritidis grown in HLA-B27- or HLA-A2-transfected cells. The microarray consisted of 5080 oligonucleotides from different serovars of Salmonella including S. Enteritidis PT4-specific genes. Bacterial RNA was isolated from the infected HLA-B27- or HLA-A2-transfected cells, reverse-transcribed to cDNA, and hybridized with the oligonucleotides on the microarrays. Some microarray results were confirmed by RT-PCR. Results When gene expression was compared between Salmonella grown in HLA-B27 cells and in HLA-A2 cells, 118 of the 4610 S. Enteritidis-related genes differed in expression at 8 h after infection, but no significant difference was detectable at 2 h after infection. These differentially expressed genes are mainly involved in Salmonella virulence, DNA replication, energy conversion and metabolism, and uptake and metabolism of nutrient substances, etc. The difference suggests HLA-B27-dependent modulation of Salmonella gene expression, resulting in increased Salmonella replication in HLA-B27-positive cells. Among the up-regulated genes were those located in Salmonella pathogenicity island (SPI-2, which play a central role in intracellular survival and replication of Salmonella. Conclusions This is the first report to show the regulation of Salmonella gene expression by HLA-B27 during infection of host cells. This regulation probably leads to increased Salmonella survival and replication in HLA-B27-positive cells. SPI-2 genes seem to contribute significantly to the increased replication.

Hinton Jay CD

2010-07-01

 
 
 
 
101

A Case of Human Leukocyte Antigen (HLA) B27-Positive Intestinal Behçet's Disease with Crohn's Disease-Like Anal Fistulas  

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A 49-year-old male was admitted to our hospital with complaints of perianal pain, bloody stool, and high-grade fever due to perianal abscess. Drainage was carried out; however, the patient’s complaints worsened, and biopsy findings of colonoscopy showed ulcerative colitis-like lesions. The patient was diagnosed as having Behçet’s disease with intestinal involvement, did not have HLA-B51, but did have HLA-B27. We describe a case of Behcet’s disease with colitis, making a differential di...

Tsuyoshi Kobashigawa; Yuki Nanke; Masakazu Takazoe; Kuniko Iihara; Hisashi Yamanaka; Shigeru Kotake

2014-01-01

102

Ankylosing spondylitis, HLA-B27, and Klebsiella: a study of lymphocyte reactivity of anti-Klebsiella sera.  

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Twenty three anti-Klebsiella antisera were tested for their cytotoxic activity and four for their binding capacity for peripheral blood lymphocytes (PBL) from patients with HLA-B27 positive ankylosing spondylitis (AS+B27+) and from B27 positive (AS-B27+) and B27 negative (AS-B27-) healthy individuals. None of the antisera showed specific activity against PBL from any particular group. The antisera tested included two anti-Klebsiella K43 sera provided by an Australian group, who have reported ...

Singh, B.; Milton, J. D.; Woodrow, J. C.

1986-01-01

103

Persistence of HLA-B27 cross-reactive bacteria in bowel flora of patients with ankylosing spondylitis.  

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Previous studies have shown that antisera raised in rabbits to certain enteric bacteria (cross-reactive bacteria) are capable of specifically lysing in a 51chromium-release lymphocytotoxicity test the lymphocytes of HLA-B27-positive (B27+) patients with ankylosing spondylitis (AS). This study investigated the clinical relevance of this finding by ascertaining whether Escherichia coli isolated from the rectal swabs of 20 B27+ AS patients (B27+ AS+) and 46 controls (35 B27- AS- and 11 B27+ AS-)...

Prendergast, J. K.; Mcguigan, L. E.; Geczy, A. F.; Kwong, T. S.; Edmonds, J. P.

1984-01-01

104

Tripeptidyl peptidase II is dispensable for the generation of both proteasome-dependent and proteasome-independent ligands of HLA-B27 and other class I molecules.  

Science.gov (United States)

A significant fraction of the HLA-B27-bound peptide repertoire is resistant to proteasome inhibitors. The possible implication of tripeptidyl peptidase II (TPPII) in generating this subset was analyzed by quantifying the surface re-expression of HLA-B*2705 after acid stripping in the presence of two TPPII inhibitors, butabindide and Ala-Ala-Phe-chloromethylketone. Neither decreased HLA-B27 re-expression under conditions in which TPPII activity was largely inhibited. This was in contrast to a significant effect of the proteasome inhibitor epoxomicin. The failure of TPPII inhibition to decrease surface re-expression was not limited to HLA-B27, since it was also observed in several HLA-B27-negative cell lines, including Mel JuSo. Actually, HLA class I re-expression in Mel JuSo cells increased as a function of butabindide concentration, which is consistent with an involvement of TPPII in destroying HLA class I ligands. Inhibition of TPPII with small interfering RNA also failed to decrease the surface expression of HLA class I molecules on 143B cells. Our results indicate that TPPII is dispensable for the generation of proteasome-dependent HLA class I ligands and, without excluding its role in producing some individual epitopes, this enzyme is not involved to any quantitatively significant extent, in generating the proteasome-independent HLA-B27-bound peptide repertoire. PMID:18286573

Marcilla, Miguel; Villasevil, Eugenia M; de Castro, José Antonio López

2008-03-01

105

PECULIARITIES OF BLOOD CYTOKINE SPECTRUM IN THE PATIENTS WITH REACTIVE ARTHRITIS DUE TO ETIOLOGY, INFLAMMATION ACTIVITY AND PRESENCE OF HLA-B27 ANTIGEN  

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Full Text Available The aim of our study was to investigate the cytokine profile due to the etiology, inflammation activity, and presence of HLA-B27 antigen in the patients with reactive arthritis. The study showed a direct dependence of IL-4, IL-6 and the TNF-? on the degree of activity of ReA with chronic pyelonephritis. The presence of HLA-B27 antigen in the patients with reactive arthritis and chronic pyelonephritis accompanied by an increasing of proinflammatory cytokines such as IL-1?, IL-6, PNP-?, IL-1R in comparison with the HLA-B27-negative patients. The results of study demonstrated that maximum serum levels of IL-6 and IFN-? in the patients with reactive arthritis that occurred on the background of acute urogenital infection were significantly higher than in the other etiological cases of reactive arthritis.

Khukhlina O. S.

2013-07-01

106

Peptide handling by HLA-B27 subtypes influences their biological behavior, association with ankylosing spondylitis and susceptibility to endoplasmic reticulum aminopeptidase 1 (ERAP1).  

Science.gov (United States)

HLA-B27 is strongly associated with ankylosing spondylitis (AS). We analyzed the relationship between structure, peptide specificity, folding, and stability of the seven major HLA-B27 subtypes to determine the role of their constitutive peptidomes in the pathogenicity of this molecule. Identification of large numbers of ligands allowed us to define the differences among subtype-bound peptidomes and to elucidate the peptide features associated with AS and molecular stability. The peptides identified only in AS-associated or high thermostability subtypes with identical A and B pockets were longer and had bulkier and more diverse C-terminal residues than those found only among non-AS-associated/lower-thermostability subtypes. Peptides sequenced from all AS-associated subtypes and not from non-AS-associated ones, thus strictly correlating with disease, were very rare. Residue 116 was critical in determining peptide binding, thermodynamic properties, and folding, thus emerging as a key feature that unified HLA-B27 biology. HLA-B27 ligands were better suited to TAP transport than their N-terminal precursors, and AS-associated subtype ligands were better than those from non-AS-associated subtypes, suggesting a particular capacity of AS-associated subtypes to bind epitopes directly produced in the cytosol. Peptides identified only from AS-associated/high-thermostability subtypes showed a higher frequency of ERAP1-resistant N-terminal residues than ligands found only in non-AS-associated/low-thermostability subtypes, reflecting a more pronounced effect of ERAP1 on the former group. Our results reveal the basis for the relationship between peptide specificity and other features of HLA-B27, provide a unified view of HLA-B27 biology and pathogenicity, and suggest a larger influence of ERAP1 polymorphism on AS-associated than non-AS-associated subtypes. PMID:25187574

García-Medel, Noel; Sanz-Bravo, Alejandro; Alvarez-Navarro, Carlos; Gómez-Molina, Patricia; Barnea, Eilon; Marcilla, Miguel; Admon, Arie; de Castro, José A López

2014-12-01

107

HLA-B27 Homodimers and Free H Chains Are Stronger Ligands for Leukocyte Ig-like Receptor B2 than Classical HLA Class 1  

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Possession of HLA-B27 (B27), strongly predisposes to the development of spondyloarthritis. B27 forms classical heterotrimeric complexes with beta-2-microglobulin (?2m) and peptide, and (?2m–free) free H chain (FHC) forms including B27 dimers (termed B272) at the cell surface. In this study we characterise the interaction of HLA-B27 with LILR, leukocyte Ig-like receptor (LILR)B1 and LILRB2 biophysically, biochemically and by FACS staining. LILRB1 bound to B27 heterotrimers with a KD of ...

Giles, Joanna; Shaw, Jackie; Piper, Christopher; Wong-baeza, Isabel; Mchugh, Kirsty; Ridley, Anna; Li, Demin; Lenart, Izabela; Antoniou, Antony N.; Digleria, Katilin; Kuroki, Kimiko; Maenaka, Katsumi; Bowness, Paul; Kollnberger, Simon

2012-01-01

108

HLA-B27 homodimers and free H chains are stronger ligands for leukocyte Ig-like receptor B2 than classical HLA class I.  

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Possession of HLA-B27 (B27) strongly predisposes to the development of spondyloarthritis. B27 forms classical heterotrimeric complexes with ?(2)-microglobulin (?2m) and peptide and (?2m free) free H chain (FHC) forms including B27 dimers (termed B27(2)) at the cell surface. In this study, we characterize the interaction of HLA-B27 with LILR, leukocyte Ig-like receptor (LILR)B1 and LILRB2 immune receptors biophysically, biochemically, and by FACS staining. LILRB1 bound to B27 heterotrimers ...

Giles, J.; Shaw, J.; Piper, C.; Wong-baeza, I.; Mchugh, K.; Ridley, A.; Li, D.; Lenart, I.; Antoniou, An; Digleria, K.; Kuroki, K.; Maenaka, K.; Bowness, P.; Kollnberger, S.

2012-01-01

109

Increase of HLA-DRB1*0408 and -DQB1*0301 in HLA-B27 positive reactive arthritis  

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OBJECTIVE—To study HLA class II association in reactive arthritis.?METHODS—63 patients with reactive arth-ritis and 46 with rheumatoid arthritis were included in the study. HLA-DR alleles were determined by using a sequence specific PCR method. Oligonucleotide hybridisation was used for definition of DRB1*04 subtypes and DQB1 alleles. HLA-B27 was determined by standard microcytotoxity test or by PCR. HLA-B27 subtyping was made by sequencing.?RESULTS—46 (73%) of 63 patients with re...

Tuokko, J.; Reijonen, H.; Ilonen, J.; Anttila, K.; Nikkari, S.; Mottonen, T.; Yli-kerttula, U.; Toivanen, A.

1997-01-01

110

Th17 cells expressing KIR3DL2+ and responsive to HLA-B27 homodimers are increased in Ankylosing Spondylitis  

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CD4 helper T cells producing the pro-inflammatory cytokine IL17 (Th17) have been implicated in a number of inflammatory arthritides including the Spondyloarthritides. Th17 development is promoted by IL23. Ankylosing Spondylitis (AS), the commonest Spondyloarthritis, is genetically associated with both HLA-B27 (B27) and with IL23 receptor polymorphisms, however the link remains unexplained. We have previously shown that B27 can form heavy chain dimers (termed B272), which, unlike classical HLA...

Bowness, P.; Ridley, A.; Shaw, J.; Chan, At; Wong-baeza, I.; Fleming, M.; Cummings, F.; Mcmichael, A.; Kollnberger, S.

2011-01-01

111

Invasion by Salmonella typhimurium Induces Increased Expression of the LMP, MECL, and PA28 Proteasome Genes and Changes in the Peptide Repertoire of HLA-B27  

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We have analyzed proteasomal adaptation and associated changes in the B27-bound peptide repertoire in response to cellular invasion with Salmonella. The peptide repertoire of HLA-B27 complexes was analyzed by two different methods: (i) high-pressure liquid chromatography (HPLC) profiles of newly synthesized peptides eluted from B27 following metabolic labeling with arginine and (ii) reactivities with two B27 monoclonal antibodies, Ye-2 and B27.M2, sensitive to peptide-induced conformational c...

Maksymowych, Walter P.; Ikawa, Takashi; Yamaguchi, Akihiro; Ikeda, Makoto; Mcdonald, Darrin; Laouar, Leila; Lahesmaa, Riitta; Tamura, Naoto; Khuong, Arranny; Yu, David T. Y.; Kane, Kevin P.

1998-01-01

112

Distribution of HLA-B27 subtypes in patients with ankylosing spondylitis: the disease is associated with a common determinant of the various B27 molecules.  

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HLA-B27 subtypes can be defined by cellular, serological, and biochemical techniques. The seven subtypes so far identified represent structural variants of B27 with limited variations in the amino acid sequence of the B27 molecule. The routinely typed B27 'antigen' remains a common (shared, public) determinant present on various B27 molecules. The distribution of the subtypes varies strongly among different ethnic groups and they occur in different linkage disequilibria. In the healthy Dutch ...

Breur-vriesendorp, B. S.; Dekker-saeys, A. J.; Ivanyi, P.

1987-01-01

113

Dominant role of the ERAP1 polymorphism R528K in shaping the HLA-B27 peptidome through differential processing determined by multiple peptide residues.  

Science.gov (United States)

Objective. To characterize the alterations, and their mechanism, induced in the HLA-B27-bound peptidome expressed in live cells by the natural ERAP1 polymorphisms predisposing to ankylosing spondylitis: R528K and N575D/Q725R. Methods. HLA-B*27:05-bound peptides were isolated from 3 human lymphoid cell lines expressing distinct ERAP1 variants differing at residues 528 and/or 575/725. The HPLC-fractionated peptide pools were compared by mass spectrometry based on identity in molecular mass and chromatographic retention time. The relative amounts of each shared peptide in any given cell line pair were estimated from the respective ion peak intensities. Peptide sequencing was also carried out by mass spectrometry. Results. HLA-B27-bound ligands predominant in the context of the ERAP1 variant with K528 collectively showed higher molecular mass, higher frequency of N-terminal residues resistant to ERAP1 and bulkier residues downstream the N-terminus, relative to peptides predominant in the R528 context. None of these differences were observed with ERAP1 variants differing at positions 575/725, but not at residue 528. Neither R528K nor N575D/Q725R altered the mean length of B*27:05-bound ligands. Conclusion. The R528K, but not the N575D/Q725R polymorphism, alters the expression levels of many HLA-B*27:05-bound peptides, depending on the susceptibility of their N-terminal residues to trimming and the size of amino acid side chains at multiple positions downstream the N-terminus. The significant alterations in the B*27:05 peptidome and the structural features of the peptides that determine their differential expression in distinct ERAP1 contexts can explain the association of the R528K polymorphism with ankylosing spondylitis. This article is protected by copyright. All rights reserved. PMID:25469497

Sanz-Bravo, Alejandro; Campos, José; Mazariegos, Marina S; LópezdeCastro, José A

2014-12-01

114

Transgenic hepatocarcinogenesis in the rat.  

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Although transgenic hepatocarcinogenesis has been accomplished in the mouse with a number of genetic constructs targeting the oncogene to expression primarily in the liver, no example of this process has yet been developed in the rat. Because our understanding of the multistage nature of hepatocarcinogenesis is most advanced in the rat, we have developed a strain of transgenic rats carrying the promoter-enhancer sequences of the mouse albumin gene linked 5' to the simian virus-40 T antigen ge...

Hully, J. R.; Su, Y.; Lohse, J. K.; Griep, A. E.; Sattler, C. A.; Haas, M. J.; Dragan, Y.; Peterson, J.; Neveu, M.; Pitot, H. C.

1994-01-01

115

Superior oblique tendon (Brown’s syndrome as the presenting finding in childhood onset HLA-B27-related enthesitis and juvenile idiopathic oligoarticular arthritis  

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Full Text Available We report two patients who presented with Brown’s syndrome. The first is a 7-year-old boy who at the time of his diagnosis was also found to have enthesitis and HLA-B27 positivity. The second patient was diagnosed with bilateral Brown’s syndrome at 13 months of age. At age 7 she developed a persistent oligoarticular arthritis and unilateral anterior iritis consistent with the oligoarticular Juvenile Idiopatic Arthritis (JIA phenotype. These cases highlight ophthalmologic findings and diagnostic considerations with respect to Brown’s syndrome and associated childhood onset rheumatologic disease.

C. Pham

2014-11-01

116

Superior oblique tendon (Brown's) syndrome as the presenting finding in childhood onset HLA-B27-related enthesitis and juvenile idiopathic oligoarticular arthritis.  

Science.gov (United States)

We report two patients who presented with Brown's syndrome. The first is a 7-year-old boy who at the time of his diagnosis was also found to have enthesitis and HLA-B27 positivity. The second patient was diagnosed with bilateral Brown's syndrome at 13 months of age. At age 7 she developed a persistent oligoarticular arthritis and unilateral anterior iritis consistent with the oligoarticular Juvenile Idiopatic Arthritis (JIA) phenotype. These cases highlight ophthalmologic findings and diagnostic considerations with respect to Brown's syndrome and associated childhood onset rheumatologic disease. PMID:25376959

Pham, C; Utz, V; Marcotty, A; Zeft, A; Rychwalski, P

2014-01-01

117

Raised incidence of ankylosing spondylitis among Inuit populations could be due to high HLA-B27 association and starch consumption.  

Science.gov (United States)

Ankylosing spondylitis (AS) is a chronic inflammatory arthritis mainly affecting the spinal joints. It would appear that the most likely causative agent in the development of AS is an environmental factor in the genetically susceptible, HLA-B27 positive, individuals. Extensive data from several countries support the notion that Klebsiella pneumonia bacteria are the most likely culprit in the causation of AS. These microbes possess antigens which resemble HLA-B27 and spinal collagens. Increased intake of high-starch diet is directly proportional to the gut-associated bacterial load, especially in the large intestine, and among these microbial agents, Klebsiella is considered as one of the main constituting components. Therefore, a low-starch diet intake alongside the currently used medical therapeutic modalities could be beneficial in the management of patients with early AS. It is suggested that a change in the dietary habits from high protein, low-starch marine components to the Westernized high-starch diet among the Inuit peoples of Alaska and Canada could be considered as one of the main contributing factors in the increased prevalence of AS during the last few decades within this genetically unmixed native population. PMID:25385438

Rashid, Taha; Wilson, Clyde; Ebringer, Alan

2014-11-11

118

Polymorphisms of HLA-A, -B, -Cw and DRB1 antigens in Moroccan patients with ankylosing spondylitis and a comparison of clinical features with frequencies of HLA-B*27.  

Science.gov (United States)

Ankylosing spondylitis (AS) is very often associated with human leukocyte antigen (HLA), particularly HLA-B*27. However, the strength of this association and clinical features may vary in different ethnic groups. Our study aims to assess the distribution of HLA-A, -B, -Cw and DRB1 alleles in Moroccan patients with AS and to compare the clinical features of AS and the frequencies of HLA-B27 in patients from Morocco with other series. Seventy-five patients diagnosed with AS and assessed for clinical manifestations were selected and compared to 100 healthy controls. HLA class I and II antigens were typed by polymerase chain reaction sequence-specific oligonucleotide. HLA-B27 subtypes were studied by polymerase chain reaction amplification with sequence-specific primers. HLA-B27 was found in 64% of patients. It was positively associated with younger age at disease onset, family history, and uveitis while it had a negative association with late onset. Six B*27 subtypes were identified in the AS group. HLA-B*2705 and B*2702 were the most common observed subtypes. Among other HLA genes, a significant increase in the prevalence of HLA-Cw*02 and HLA-DRB*15 was found in AS patients. HLA-B27 is involved in the predisposition of AS in the Moroccan population. HLA-B*2705 and B*2702 were the predominant subtypes supporting previous reports in Caucasian spondyloarthropathies. Other HLA genes, HLA-Cw*02 and HLA-DRB1*15, seem to confer predisposing effect to the disease. However, the lower frequency of HLA-B27 compared to the literature in our study suggests the existence of different genetic and/or environmental factors in Morocco. PMID:25626601

El Mouraghi, I; Ouarour, A; Ghozlani, I; Collantes, E; Solana, R; El Maghraoui, A

2015-02-01

119

HLA-B27 Homodimers and Free H Chains Are Stronger Ligands for Leukocyte Ig-like Receptor B2 than Classical HLA Class 1  

Science.gov (United States)

Possession of HLA-B27 (B27), strongly predisposes to the development of spondyloarthritis. B27 forms classical heterotrimeric complexes with beta-2-microglobulin (?2m) and peptide, and (?2m–free) free H chain (FHC) forms including B27 dimers (termed B272) at the cell surface. In this study we characterise the interaction of HLA-B27 with LILR, leukocyte Ig-like receptor (LILR)B1 and LILRB2 biophysically, biochemically and by FACS staining. LILRB1 bound to B27 heterotrimers with a KD of 5.3 ±1.5 ?M but did not bind B27 FHC. LILRB2 bound to B272 and B27 FHC and B27 heterotrimers with KDs of 2.5, 2.6 and 22 ±6?M respectively. Domain exchange experiments showed that B272 bound to the two membrane distal Ig-like domains of LILRB2. In FACS staining experiments, B27 dimer protein and tetramers stained LILRB2 transfectants five times more strongly than B27 heterotrimers. Moreover, LILRB2Fc bound to dimeric and other B27 FHC forms on B27-expressing cell lines more strongly than other HLA-class I FHCs. B27 transfected cells expressing B27 dimers and FHC inhibited IL-2 production by LILRB2-expressing reporter cells to a greater extent than control HLA-class I transfectants. B27 heterotrimers complexed with the L6M variant of the GAG KK10 epitope bound with a similar affinity to complexes with the wild-type KK10 epitope (with KDs of 15.0±0.8 ?M and 16.0±2.0 ?M respectively). Disulfide-dependent B27 H chain dimers and multimers are stronger ligands for LILRB2 than HLA-class I heterotrimers and H chains. The stronger interaction of B27 dimers and FHC forms with LILRB2 compared with other HLA class I could play a role in spondyloarthritis pathogenesis. PMID:22593621

Giles, Joanna; Shaw, Jackie; Piper, Christopher; Wong-Baeza, Isabel; McHugh, Kirsty; Ridley, Anna; Li, Demin; Lenart, Izabela; Antoniou, Antony N.; DiGleria, Katilin; Kuroki, Kimiko; Maenaka, Katsumi; Bowness, Paul; Kollnberger, Simon

2013-01-01

120

HLA-B27 Test  

Science.gov (United States)

... diseases such as ankylosing spondylitis (AS) , juvenile rheumatoid arthritis (JRA) , reactive arthritis (of which one subset is Reiter syndrome), and ... confirm a suspected diagnosis of ankylosing spondylitis (AS) , reactive arthritis , juvenile rheumatoid arthritis (JRA) , or sometimes anterior uveitis . ...

 
 
 
 
121

Peptide-binding motifs associated with MHC molecules common in Chinese rhesus macaques are analogous to those of human HLA supertypes and include HLA-B27-like alleles  

DEFF Research Database (Denmark)

Chinese rhesus macaques are of particular interest in simian immunodeficiency virus/human immunodeficiency virus (SIV/HIV) research as these animals have prolonged kinetics of disease progression to acquired immunodeficiency syndrome (AIDS), compared to their Indian counterparts, suggesting that they may be a better model for HIV. Nevertheless, the specific mechanism(s) accounting for these kinetics remains unclear. The study of major histocompatibility complex (MHC) molecules, including their MHC/peptide-binding motifs, provides valuable information for measuring cellular immune responses and deciphering outcomes of infection and vaccine efficacy. In this study, we have provided detailed characterization of six prevalent Chinese rhesus macaque MHC class I alleles, yielding a combined phenotypic frequency of 29 %. The peptide-binding specificity of two of these alleles, Mamu-A2*01:02 and Mamu-B*010:01, as well as the previously characterized allele Mamu-B*003:01 (and Indian rhesus Mamu-B*003:01), was found tobe analogous to that of alleles in the HLA-B27 supertype family. Specific alleles in the HLA-B27 supertype family, including HLA-B*27:05, have been associated with long-term nonprogression to AIDS in humans. All six alleles characterized in the present study were found to have specificities analogous to HLA supertype alleles. These data contribute to the concept that Chinese rhesus macaque MHC immunogenetics is more similar to HLA than their Indian rhesus macaque counterparts and thereby warrants further studies to decipher the role of these alleles in the context of SIV infection.

Mothé, Bianca R.; Southwood, Scott

2013-01-01

122

KIR3DL2 binds to HLA-B27 dimers and free heavy chains more strongly than other HLA class I and promotes the expansion of T cells in ankylosing spondylitis  

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The Human Leukocyte Antigen HLA-B27(B27) is strongly associated with the spondyloarthritides. B27 can be expressed at the cell surface of antigen presenting cells (APC) as both classical ?2m-associated B27 and as B27 free heavy chain forms (FHC) including disulphide-bonded heavy chain homodimers (termed B272). B27 FHC forms but not classical B27 bind to KIR3DL2. HLA-A3 which is not associated with spondyloarthritis (SpA) is also a ligand for KIR3DL2. Here we show that B272 and B27 FHC bind m...

Wong-baeza, Isabel; Ridley, Anna; Shaw, Jackie; Hatano, Hiroko; Rysnik, Oliwia; Mchugh, Kirsty; Piper, Christopher; Brackenbridge, Simon; Fernandes, Ricardo; Chan, Anthoni; Bowness, Paul; Kollnberger, Simon

2013-01-01

123

KIR3DL2 binds to HLA-B27 dimers and free H chains more strongly than other HLA class I and promotes the expansion of T cells in ankylosing spondylitis.  

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The human leukocyte Ag HLA-B27 (B27) is strongly associated with the spondyloarthritides. B27 can be expressed at the cell surface of APC as both classical [beta]2-microglobulin-associated B27 and B27 free H chain forms (FHC), including disulfide-bonded H chain homodimers (termed B272). B27 FHC forms, but not classical B27, bind to KIR3DL2. HLA-A3, which is not associated with spondyloarthritis (SpA), is also a ligand for KIR3DL2. In this study, we show that B272 and B27 FHC bind more strongl...

Wong-baeza, I.; Ridley, A.; Shaw, J.; Hatano, H.; Rysnik, O.; Mchugh, K.; Piper, C.; Brackenbridge, S.; Fernandes, R.; Chan, A.; Bowness, P.; Kollnberger, S.

2013-01-01

124

Focal cerebral ischemia in the TNFalpha-transgenic rat  

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Abstract Background To determine if chronic elevation of the inflammatory cytokine, tumor necrosis factor-? (TNF?), will affect infarct volume or cortical perfusion after focal cerebral ischemia. Methods Transgenic (TNF?-Tg) rats overexpressing the murine TNF? gene in brain were prepared by injection of mouse DNA into rat oocytes. Brain levels of TNF? mRNA and protein were measured and compared between TNF?-Tg and non-transgenic (non-Tg) littermates. Mean ...

Springer Joe E; Scheff Stephen; Kindy Mark S; Creed, Pettigrew L.; Kryscio Richard J; Li Yizhao; Grass David S

2008-01-01

125

A transgenic rat with ubiquitous expression of firefly luciferase gene  

Science.gov (United States)

In vivo imaging strategies provide cellular and molecular events in real time that helps us to understand biological processes in living animals. The development of molecular tags such as green fluorescent proteins and luciferase from the firefly Photinus pyralis has lead to a revolution in the visualization of complex biochemical processes. We developed a novel inbred transgenic rat strain containing firefly luciferase based on the transgenic (Tg) technique in rats. This Tg rat expressed the luciferase gene ubiquitously under control of the ROSA26 promoter. Cellular immune responsiveness against the luciferase protein was evaluated using conventional skin grafting and resulted in the long-term acceptance of Tg rat skin on wild-type rats. Strikingly, organ transplant with heart and small bowel demonstrated organ viability and graft survival, suggesting that cells from luciferase-Tg are transplantable to track their fate. Taking advantage of the less immunogenic luciferase, we also tested the role of hepatocyte-infusion in a liver injury model, and bone marrow-derived cells in a skin defect model. Employed in conjunction with modern advances in optical imaging, this luciferase-Tg rat system provides an innovative animal tool and a new means of facilitating biomedical research such as in the case of regeneration medicine.

Hakamata, Yoji; Murakami, Takashi; Kobayashi, Eiji

2006-02-01

126

A preliminary exploration on DNA methylation of transgene across generations in transgenic rats  

Science.gov (United States)

Epigenetic heritability is an important issue in the field of genetics and also in the development of many human diseases. In this study, we created a transgenic rat model and investigated the transgenerational methylation patterns in these animals. The transgene DNA fragment was unmethylated before it was injected into the pronucleus, so it is a good model to study the inheritance of DNA methylation patterns. We performed bisulfite sequencing on 23 CpG dinucleotides on the transgene across three generations in two tissues. We observed that the transgene was heavily methylated in the liver (87.53%) from the founder generation, whereas its methylation rate was much lower in the kidney (70.47%). Spearman correlation analysis showed that there was a strong correlation on the methylation status between different generations in the same tissue, which was observed in both liver and kidney, and among all individuals in this pedigree. This study provided some evidence that DNA methylation patterns acquired in the founder animal can be passed to the offspring. PMID:25659774

Li, Qiling; Xu, Wei; Cui, Ye; Ma, Li; Richards, Jendai; Li, Wenzhi; Ma, Yamin; Fu, Guoxing; Bythwood, Tameka; Wang, Yueling; Li, Xu; Song, Qing

2015-01-01

127

HIV-1 transgenic rats develop T cell abnormalities  

International Nuclear Information System (INIS)

HIV-1 infection leads to impaired antigen-specific T cell proliferation, increased susceptibility of T cells to apoptosis, progressive impairment of T-helper 1 (Th1) responses, and altered maturation of HIV-1-specific memory cells. We have identified similar impairments in HIV-1 transgenic (Tg) rats. Tg rats developed an absolute reduction in CD4+ and CD8+ T cells able to produce IFN-? following activation and an increased susceptibility of T cells to activation-induced apoptosis. CD4+ and CD8+ effector/memory (CD45RC-CD62L-) pools were significantly smaller in Tg rats compared to non-Tg controls, although the converse was true for the naieve (CD45RC+CD62L+) T cell pool. Our interpretation is that the HIV transgene causes defects in the development of T cell effector function and generation of specific effector/memory T cell subsets, and that activation-induced apoptosis may be an essential factor in this process

128

Developing tTA Transgenic Rats for Inducible and Reversible Gene Expression  

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To develop transgenic lines for conditional expression of desired genes in rats, we generated several lines of the transgenic rats carrying the tetracycline-controlled transactivator (tTA) gene. Using a vigorous, ubiquitous promoter to drive the tTA transgene, we obtained widespread expression of tTA in various tissues. Expression of tTA was sufficient to strongly activate its reporter gene, but was below the toxicity threshold. We examined the dynamics of Doxycycline (Dox)-regulated gene ...

Hongxia Zhou, Cao Huang

2009-01-01

129

Regulatory regions of rat insulin I gene necessary for expression in transgenic mice.  

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Ten transgenic mouse lines harboring the -346/-103 fragment of the rat insulin I enhancer linked to a heterologous promoter and a reporter gene (Eins-Ptk-CAT construct) were produced. Expression of the hybrid transgene was essentially observed in pancreas and to a lesser extent in brain. These results indicate that the rat insulin I promoter is dispensable for pancreatic expression. This insulin gene sequence is the shortest fragment described as conferring tissue-specific expression in trans...

Dandoy-dron, F.; Monthioux, E.; Jami, J.; Bucchini, D.

1991-01-01

130

Protection against hyperacute xenograft rejection of transgenic rat hearts expressing human decay accelerating factor (DAF) transplanted into primates.  

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BACKGROUND: Production of transgenic pigs for multiple transgenes is part of a potential strategy to prevent immunological events involved in xenograft rejection. Use of a genetically engineerable rodent as a donor in primates could allow testing in vivo of the effects of different transgenes on controlling xenograft rejection. As a first step in the development of a donor containing multiple transgenes, transgenic rats for human decay-accelerating factor (DAF) were used as heart donors to te...

Charreau, B.; Ma?©noret, S.; Tesson, L.; Azimzadeh, A.; Audet, M.; Wolf, P.; Marquet, R.; Verbakel, C.; Ijzermans, J.; Cowan, P.; Pearse, M.; D Apice, A.; Soulillou, J. P.; Anegon, I.

1999-01-01

131

A transgenic rat expressing human APP with the Swedish Alzheimer's disease mutation  

DEFF Research Database (Denmark)

In recent years, transgenic mice have become valuable tools for studying mechanisms of Alzheimer's disease (AD). With the aim of developing an animal model better for memory and neurobehavioural testing, we have generated a transgenic rat model of AD. These animals express human amyloid precursor protein (APP) containing the Swedish AD mutation. The highest level of expression in the brain is found in the cortex, hippocampus, and cerebellum. Starting after the age of 15 months, the rats show increased tau phosphorylation and extracellular Abeta staining. The Abeta is found predominantly in cerebrovascular blood vessels with very rare diffuse plaques. We believe that crossing these animals with mutant PS1 transgenic rats will result in accelerated plaque formation similar to that seen in transgenic mice.

Folkesson, Ronnie; Malkiewicz, Katarzyna

2007-01-01

132

Establishment of an Invasive Prostate Cancer Model in Transgenic Rats by Intermittent Testosterone Administration  

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We have established a transgenic rat for adenocarcinoma of the prostate (TRAP) model that features uniform adenocarcinoma development in prostatic lobes at high incidence within a short experimental period. However, no invasive carcinomas with reactive stroma characteristics similar to those in man were observed. We therefore have focused on a new model for invasive carcinoma of the prostate using TRAP rats. In experiment 1, male TRAP rats in groups 1 and 2 were treated with orchiectomy at da...

Sato, Shinya; Suzuki, Shugo; Naiki-ito, Aya; Komiya, Masami; Ne, Long; Kato, Hiroyuki; Sagawa, Hiroyuki; Yamashita, Yoriko; Shirai, Tomoyuki; Takahashi, Satoru

2014-01-01

133

Reduced sodium-proton exchange activity in lymphocytes from transgenic rats.  

Science.gov (United States)

We investigated sodium-proton (Na(+)-H+) exchange activity in transgenic TGR(mRen-2)27 rats, a strain showing fulminant hypertension after the mouse Ren-2d renin gene has been integrated into its genome, in age-matched normotensive Sprague-Dawley (SD) rats, in spontaneously hypertensive rats (SHR) from the Münster strain, and in normotensive Wistar-Kyoto (WKY) rats. From each strain Na(+)-H+ exchange activity was determined in lymphocytes using the pH-sensitive fluorescent dye 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein acetoxymethyl ester (BCECF-AM) by measuring the recovery rate of cytosolic pH (pHi) after intracellular acidification. Resting pHi was not significantly different in transgenic rats (n = 10) compared with SD rats (n = 10) (7.305 +/- 0.038 versus 7.337 +/- 0.031; mean +/- SEM), but resting pHi was significantly lower in lymphocytes from SHR (n = 12) compared with their normotensive WKY counterparts (n = 12) (7.232 +/- 0.030 versus 7.377 +/- 0.022; P dpHi/s; P dpHi/s; P < .05). The apparent half-maximal activation of Na(+)-H+ exchange was not significantly different in the strains tested. The present study indicates that hypertension in transgenic rats is not related to Na(+)-H+ exchange overactivity. PMID:8082942

Tepel, M; Klaus, T; Laukemper, S; Zidek, W

1994-09-01

134

Microstructural changes observed with DKI in a transgenic Huntington rat model: evidence for abnormal neurodevelopment.  

Science.gov (United States)

Huntington Disease (HD) is a fatal neurodegenerative disorder, caused by a mutation in the Huntington gene. Although HD is most often diagnosed in mid-life, the key to its clinical expression may be found during brain maturation. In the present work, we performed in vivo diffusion kurtosis imaging (DKI) in order to study brain microstructure alterations in developing transgenic HD rat pups. Several developing brain regions, relevant for HD pathology (caudate putamen, cortex, corpus callosum, external capsule and anterior commissure anterior), were examined at postnatal days 15 (P15) and 30 (P30), and DKI results were validated with histology. At P15, we observed higher mean (MD) and radial (RD) diffusivity values in the cortex of transgenic HD rat pups. In addition, at the age of P30, lower axial kurtosis (AK) values in the caudate putamen of transgenic HD pups were found. At the level of the external capsule, higher MD values at P15 but lower MD and AD values at P30 were detected. The observed DKI results have been confirmed by myelin basic protein immunohistochemistry, which revealed a reduced fiber staining as well as less ordered fibers in transgenic HD rat pups. These results indicate that neuronal development in young transgenic HD rat pups occurs differently compared to controls and that the presence of mutant huntingtin has an influence on postnatal brain development. In this context, various diffusivity parameters estimated by the DKI model are a powerful tool to assess changes in tissue microstructure and detect developmental changes in young transgenic HD rat pups. PMID:21906685

Blockx, Ines; De Groof, Geert; Verhoye, Marleen; Van Audekerke, Johan; Raber, Kerstin; Poot, Dirk; Sijbers, Jan; Osmand, Alexander P; Von Hörsten, Stephan; Van der Linden, Annemie

2012-01-16

135

Tauroursodeoxycholic acid prevents retinal degeneration in transgenic P23H rats  

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Purpose. To evaluate the preventive effect of tauroursodeoxycholic acid (TUDCA) on photoreceptor degeneration, synaptic connectivity and functional activity of the retina in the transgenic P23H rat, an animal model of autosomal dominant retinitis pigmentosa (RP). Methods. P23H line-3 rats were injected with TUDCA once a week from postnatal day (P)21 to P120, in parallel with vehicle-administered controls. At P120, functional activity of the retina was evaluated by electroretinographic (ERG) r...

Ferna?ndez Sa?nchez, Laura; Lax Zapata, Pedro; Pinilla Lozano, Isabel; Marti?n Nieto, Jose?; Cuenca Navarro, Nicola?s

2011-01-01

136

Cognitive impairment in the Tg6590 transgenic rat model of Alzheimer's disease  

DEFF Research Database (Denmark)

Recently, interest in the rat as an animal model of Alzheimer's disease (AD) has been growing. We have previously described the Tg6590 transgenic rat line expressing the amyloid precursor protein containing the Swedish AD mutation (K670M/N671L) that shows early stages of Abeta deposition, predominantly in cerebrovascular blood vessels, after 15 months of age. Here we show that by the age of 9 months, that is long before the appearance of Abeta deposits, the Tg6590 rats exhibit deficits in the Morris water maze spatial navigation task and altered spontaneous behaviour in the open-field test. The levels of soluble Abeta were elevated both in the hippocampus and cortex of transgenic animals. Magnetic resonance imaging showed no major changes in the brains of transgenic animals, although they tended to have enlarged lateral ventricles when compared to control animals. The Tg6590 transgenic rat line should prove a suitable model of early AD for advanced studies including serial cerebrospinal fluid sampling, electrophysiology, neuroimaging or complex behavioural testing.

Kloskowska, Ewa; Pham, Therese M

2010-01-01

137

Specific expression of an oxytocin-enhanced cyan fluorescent protein fusion transgene in the rat hypothalamus and posterior pituitary  

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We have generated rats bearing an oxytocin (OXT)-enhanced cyan fluorescent protein (eCFP) fusion transgene designed from a murine construct previously shown to be faithfully expressed in transgenic mice. In situ hybridisation histochemistry revealed that the OXT-eCFP fusion gene was expressed in the supraoptic (SON) and the paraventricular nuclei (PVN) in these rats. The fluorescence emanating from eCFP was observed only in the SON, the PVN, the internal layer of the median eminence (ME) and ...

Katoh, Akiko; Fujihara, Hiroaki; Ohbuchi, Toyoaki; Onaka, Tatsushi; Young, W. Scott; Dayanithi, Govindan; Yamasaki, Yuka; Kawata, Mitsuhiro; Suzuki, Hitoshi; Otsubo, Hiroki; Suzuki, Hideaki; Murphy, David; Ueta, Yoichi

2010-01-01

138

HIV-1 transgene expression in rats causes oxidant stress and alveolar epithelial barrier dysfunction  

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Abstract Background HIV-infected individuals are at increased risk for acute and chronic airway disease even though there is no evidence that the virus can infect the lung epithelium. Although HIV-related proteins including gp120 and Tat can directly cause oxidant stress and cellular dysfunction, their effects in the lung are unknown. The goal of this study was to determine the effects of HIV-1 transgene expression in rats on alveolar epithelial barrier function. Alveolar epi...

Jacob Barbara A; Joshi Pratibha C; Fan Xian; Lassiter Coy; Sutliff Roy L; Jones Dean P; Koval Michael; Guidot David M

2009-01-01

139

Meloxicam Blocks Neuroinflammation, but Not Depressive-Like Behaviors, in HIV-1 Transgenic Female Rats  

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Adolescents living with human immunodeficiency virus (HIV) comprise approximately 12% of the HIV-positive population worldwide. HIV-positive adolescents experience a higher rate of clinical depression, a greater risk of sexual and drug abuse behaviors, and a decreased adherence to highly active antiretroviral therapies (HAART). Using adolescent HIV-1 transgenic rats (HIV-1 tg) that display related immune response alterations and pathologies, this study tested the hypothesis that developmental...

Nemeth, Christina L.; Glasper, Erica R.; Harrell, Constance S.; Malviya, Sanjana A.; Otis, Jeffrey S.; Neigh, Gretchen N.

2014-01-01

140

Alterations in the immuno-skeletal interface drive bone destruction in HIV-1 transgenic rats  

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Osteoporosis and bone fractures are increasingly recognized complications of HIV-1 infection. Although antiretroviral therapy itself has complex effects on bone turnover, it is now evident that the majority of HIV-infected individuals already exhibit reduced bone mineral density before therapy. The mechanisms responsible are likely multifactorial and have been difficult to delineate in humans. The HIV-1 transgenic rat recapitulates many key features of human AIDS. We now demonstrate that, lik...

Vikulina, Tatyana; Fan, Xian; Yamaguchi, Masayoshi; Roser-page, Susanne; Zayzafoon, Majd; Guidot, David M.; Ofotokun, Ighovwerha; Weitzmann, M. Neale

2010-01-01

 
 
 
 
141

Derivation and Characterization of Embryonic Stem Cells Lines Derived from Transgenic Fischer 344 and Dark Agouti Rats  

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Rat embryonic stem cell (ESC) lines are not widely available, and there are only 2 lines available for distribution. Here, ESC lines were derived and characterized from Fischer 344 (F344) rats that express marker transgenes either ?-galactosidase or human placental alkaline phosphatase (AP), nontransgenic F344 rats, and from Dark Agouti (DA) rats. The ESC lines were maintained in an undifferentiated state as characterized by colony morphology, expression of Oct4, Nanog, Sox-2, Cdx2, and Stel...

Hong, James; He, Hong; Weiss, Mark L.

2012-01-01

142

Chronic alcohol ingestion exacerbates skeletal muscle myopathy in HIV-1 transgenic rats  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Separately, chronic alcohol ingestion and HIV-1 infection are associated with severe skeletal muscle derangements, including atrophy and wasting, weakness, and fatigue. One prospective cohort study reported that 41% of HIV-infected patients met the criteria for alcoholism, however; few reports exist on the co-morbid effects of these two disease processes on skeletal muscle homeostasis. Thus, we analyzed the atrophic effects of chronic alcohol ingestion in HIV-1 transgenic rats and identified alterations to several catabolic and anabolic factors. Findings Relative plantaris mass, total protein content, and fiber cross-sectional area were reduced in each experimental group compared to healthy, control-fed rats. Alcohol abuse further reduced plantaris fiber area in HIV-1 transgenic rats. Consistent with previous reports, gene levels of myostatin and its receptor activin IIB were not increased in HIV-1 transgenic rat muscle. However, myostatin and activin IIB were induced in healthy and HIV-1 transgenic rats fed alcohol for 12 weeks. Catabolic signaling factors such as TGF?1, TNF?, and phospho-p38/total-p38 were increased in all groups compared to controls. There was no effect on IL-6, leukemia inhibitory factor (LIF, cardiotrophin-1 (CT-1, or ciliary neurotrophic factor (CNTF in control-fed, transgenic rats. However, the co-morbidity of chronic alcohol abuse and HIV-1-related protein expression decreased expression of the two anabolic factors, CT-1 and CNTF. Conclusions Consistent with previous reports, alcohol abuse accentuated skeletal muscle atrophy in an animal model of HIV/AIDS. While some catabolic pathways known to drive alcoholic or HIV-1-associated myopathies were also elevated in this co-morbid model (e.g., TGF?1, consistent expression patterns were not apparent. Thus, specific alterations to signaling mechanisms such as the induction of the myostatin/activin IIB system or reductions in growth factor signaling via CT-1- and CNTF-dependent mechanisms may play larger roles in the regulation of muscle mass in alcoholic, HIV-1 models.

Bratina Margaux A

2011-08-01

143

In vivo cell kinetics of the bone marrow transplantation using dual colored transgenic rat system  

Science.gov (United States)

Because bone marrow is an adequate site for bone marrow stem cells, intra-bone marrow - bone marrow transplantation (IBM-BMT) is an efficient strategy for bone marrow transplantation (BMT). However, the fate of the transplanted cells remains unclear. Herein, we established a dual-colored transgenic rat system utilizing green fluorescent protein (GFP) and a luciferase (luc) marker. We then utilized this system to investigate the in vivo kinetics of transplanted bone marrow cells (BMCs) after authentic intravenous (IV)-BMT or IBM-BMT. The in vivo fate of the transplanted cells was tracked using an in vivo luminescent imaging technique; alterations in peripheral blood chimerism were also followed using flow cytometry. IBM-BMT and IV-BMT were performed using syngeneic and allogeneic rat combinations. While no difference in the proliferation pattern was observed between the two treatment groups at 7 days after BMT, different distribution patterns were clearly observed during the early phase. In the IBM-BMT-treated rats, the transplanted BMCs were engrafted immediately at the site of the injected bone marrow and expanded more rapidly than in the IV-BMT-treated rats during this phase. Graft-versus-host disease was also visualized. Our bio-imaging system using dual-colored transgenic rats is a powerful tool for performing quantitative and morphological assessments in vivo.

Kai, Kotaro; Teraoka, Satoshi; Adachi, Yasushi; Ikehara, Susumu; Murakami, Takashi; Kobayashi, Eiji

2008-02-01

144

Bioimaging of DsRed fluorescence in the transgenic rat liver  

Science.gov (United States)

We developed the Alb-DsRed2 transgenic (Tg) rat designed with liver-specific expression of the red fluorescent protein, DsRed2. Herein, we report high expression of DsRed2 in neonate liver of both sexes, although they were sexually dimorphic and exhibited a male-specific pattern in adult rats. In an effort to examine the expression in each animal under development, we employed an in vivo Bio-imaging system to quantitatively estimate hepatic DsRed2 expression levels. The temporal profiles pertaining to DsRed expression were similar in male and female Tg rats until 28 days old. The levels in both sexes decreased gradually following birth, and were not detectable at 21 days. Subsequently, expression in males increased again at 35 days and was maintained at a persistently high level thereafter. On the other hand, expression in females disappeared steadily. Although hepatic DsRed expression levels in gonadectomized Tg rats was not significantly different, DsRed expression in hypophysectomized female Tg rats appeared dramatically 72 hr following operation. Hepatocytes were collected from adult Tg rats and cultured in conditioning medium. DsRed expression in female hepatocytes could be detected 72 hr following culturing. These results suggest that hepatic DsRed expression in female rats is regulated in vivo by the pituitary. This report is shows use of Alb-DsRed2 Tg rats in conjunction with a novel bio-imaging system represents a powerful experimental system.

Arao, Yukitomo; Hakamata, Yoji; Igarashi, Yuka; Sato, Yuki; Murakami, Takashi; Kobayashi, Eiji

2006-02-01

145

Does the microbiome play a causal role in spondyloarthritis?  

Science.gov (United States)

The purpose of this study is to review the potential causal role of the microbiome in the pathogenesis of spondyloarthritis. The method used for the study is literature review. The microbiome plays a major role in educating the immune response. The microbiome is strongly implicated in inflammatory bowel disease which has clinical and genetic overlap with spondyloarthritis. The microbiome also plays a causal role in bowel and joint disease in HLA B27/human beta 2 microglobulin transgenic rats. The mechanism(s) by which HLA B27 could influence the microbiome is unknown but theories include an immune response gene selectivity, an effect on dendritic cell function, or a mucosal immunodeficiency. Bacteria are strongly implicated in the pathogenesis of spondyloarthritis. Studies to understand how HLA B27 affects bacterial ecosystems should be encouraged. PMID:24810703

Rosenbaum, James T; Lin, Phoebe; Asquith, Mark; Costello, Mary-Ellen; Kenna, Tony J; Brown, Matthew A

2014-06-01

146

Increased blood pressure and water intake in transgenic mice expressing rat tonin in the brain.  

Science.gov (United States)

Tonin is a serine proteinase of the kallikrein family that can produce angiotensin II directly from angiotensinogen. To clarify the importance of this enzyme for central nervous control of the cardiovascular system, we generated transgenic mice, TGM(rTon), that express rat tonin in astrocytes. These mice present high levels of tonin mRNA and activity specifically in the brain. As a consequence, TGM(rTon) develop increased blood pressure and water intake. Lisinopril, an ACE inhibitor, is less hypotensive for transgenic mice than for control animals. The AT(1) receptor antagonist candesartan equally lowers blood pressure in transgenic and in control mice. Plasma angiotensin II, but not angiotensin I, is increased in TGM(rTon) compared to the wild type, suggesting release of the peptide from the brain into the circulation. However, AT(1) receptors are desensitized in this transgenic model, as demonstrated by a blunted pressor response to intravenous application of angiotensin II. In conclusion, tonin in the brain may represent an alternative pathway for angiotensin II generation with effects on the cardiovascular system. PMID:20180641

Cardoso, Cibele C; Alenina, Natalia; Ferreira, Anderson J; Qadri, Fatimunnisa; Lima, Mércia P; Gross, Volkmar; Todiras, Mihail; Pesquero, João B; Pesquero, Jorge L; Bader, Michael

2010-04-01

147

Megaesophagus in a line of transgenic rats: a model of achalasia.  

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Megaesophagus is defined as the abnormal enlargement or dilatation of the esophagus, characterized by a lack of normal contraction of the esophageal walls. This is called achalasia when associated with reduced or no relaxation of the lower esophageal sphincter (LES). To date, there are few naturally occurring models for this disease. A colony of transgenic (Pvrl3-Cre) rats presented with megaesophagus at 3 to 4 months of age; further breeding studies revealed a prevalence of 90% of transgene-positive animals having megaesophagus. Affected rats could be maintained on a total liquid diet long term and were shown to display the classic features of dilated esophagus, closed lower esophageal sphincter, and abnormal contractions on contrast radiography and fluoroscopy. Histologically, the findings of muscle degeneration, inflammation, and a reduced number of myenteric ganglia in the esophagus combined with ultrastructural lesions of muscle fiber disarray and mitochondrial changes in the striated muscle of these animals closely mimic that seen in the human condition. Muscle contractile studies looking at the response of the lower esophageal sphincter and fundus to electrical field stimulation, sodium nitroprusside, and L-nitro-L-arginine methyl ester also demonstrate the similarity between megaesophagus in the transgenic rats and patients with achalasia. No primary cause for megaesophagus was found, but the close parallel to the human form of the disease, as well as ease of care and manipulation of these rats, makes this a suitable model to better understand the etiology of achalasia as well as study new management and treatment options for this incurable condition. PMID:24457157

Pang, J; Borjeson, T M; Muthupalani, S; Ducore, R M; Carr, C A; Feng, Y; Sullivan, M P; Cristofaro, V; Luo, J; Lindstrom, J M; Fox, J G

2014-11-01

148

Specific expression of an oxytocin-enhanced cyan fluorescent protein fusion transgene in the rat hypothalamus and posterior pituitary.  

Science.gov (United States)

We have generated rats bearing an oxytocin (OXT)-enhanced cyan fluorescent protein (eCFP) fusion transgene designed from a murine construct previously shown to be faithfully expressed in transgenic mice. In situ hybridisation histochemistry revealed that the Oxt-eCfp fusion gene was expressed in the supraoptic nucleus (SON) and the paraventricular nucleus (PVN) in these rats. The fluorescence emanating from eCFP was observed only in the SON, the PVN, the internal layer of the median eminence and the posterior pituitary (PP). In in vitro preparations, freshly dissociated cells from the SON and axon terminals showed clear eCFP fluorescence. Immunohistochemistry for OXT and arginine vasopressin (AVP) revealed that the eCFP fluorescence co-localises with OXT immunofluorescence, but not with AVP immunofluorescence in the SON and the PVN. Although the expression levels of the Oxt-eCfp fusion gene in the SON and the PVN showed a wide range of variations in transgenic rats, eCFP fluorescence was markedly increased in the SON and the PVN, but decreased in the PP after chronic salt loading. The expression of the Oxt gene was significantly increased in the SON and the PVN after chronic salt loading in both non-transgenic and transgenic rats. Compared with wild-type animals, euhydrated and salt-loaded male and female transgenic rats showed no significant differences in plasma osmolality, sodium concentration and OXT and AVP levels, suggesting that the fusion gene expression did not disturb any physiological processes. These results suggest that our new transgenic rats are a valuable new tool to identify OXT-producing neurones and their terminals. PMID:20026620

Katoh, Akiko; Fujihara, Hiroaki; Ohbuchi, Toyoaki; Onaka, Tatsushi; Young, W Scott; Dayanithi, Govindan; Yamasaki, Yuka; Kawata, Mitsuhiro; Suzuki, Hitoshi; Otsubo, Hiroki; Suzuki, Hideaki; Murphy, David; Ueta, Yoichi

2010-03-01

149

Polyethylenimine-mediated expression of transgenes in the acinar cells of rats salivary glands in vivo  

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Non viral-mediated transfection of plasmid DNA provides a fast and reliable way to express various transgenes in selected cell populations in live animals. Here, we show an improvement of a previously published method that is based on injecting plasmid DNA into the ductal system of the salivary glands in live rats. Specifically, using complexes between plasmid DNA and polyethyleneimine (PEI) we show that the expression of the transgenes is directed selectively to the salivary acinar cells. PEI does not affect the ability of cells to undergo regulated exocytosis, which was one of the main drawbacks of the previous methods. Moreover PEI does not affect the proper localization and targeting of transfected proteins, as shown for the apical plasma membrane water channel aquaporin 5 (AQP5). Overall, this approach, coupled with the use of intravital microscopy, permits to conduct localization and functional studies under physiological conditions, in a rapid, reliable, and affordable fashion.

Sramkova, Monika; Parente, Laura; Wigand, Timothy; Aye, Myo-Pale'; Shitara, Akiko; Weigert, Roberto

2015-01-01

150

HIV-1 transgene expression in rats causes oxidant stress and alveolar epithelial barrier dysfunction  

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Full Text Available Abstract Background HIV-infected individuals are at increased risk for acute and chronic airway disease even though there is no evidence that the virus can infect the lung epithelium. Although HIV-related proteins including gp120 and Tat can directly cause oxidant stress and cellular dysfunction, their effects in the lung are unknown. The goal of this study was to determine the effects of HIV-1 transgene expression in rats on alveolar epithelial barrier function. Alveolar epithelial barrier function was assessed by determining lung liquid clearance in vivo and alveolar epithelial monolayer permeability in vitro. Oxidant stress in the alveolar space was determined by measuring the glutathione redox couple by high performance liquid chromatography, and the expression and membrane localization of key tight junction proteins were assessed. Finally, the direct effects of the HIV-related proteins gp120 and Tat on alveolar epithelial barrier formation and tight junction protein expression were determined. Results HIV-1 transgene expression caused oxidant stress within the alveolar space and impaired epithelial barrier function even though there was no evidence of overt inflammation within the airways. The expression and membrane localization of the tight junction proteins zonula occludens-1 and occludin were decreased in alveolar epithelial cells from HIV-1 transgenic rats. Further, treating alveolar epithelial monolayers from wild type rats in vitro with recombinant gp120 or Tat for 24 hours reproduced many of the effects on zonula occludens-1 and occludin expression and membrane localization. Conclusion Taken together, these data indicate that HIV-related proteins cause oxidant stress and alter the expression of critical tight junction proteins in the alveolar epithelium, resulting in barrier dysfunction.

Jacob Barbara A

2009-02-01

151

Retinal ganglion cell survival and axon regeneration in WldS transgenic rats after optic nerve crush and lens injury  

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Full Text Available Abstract Background We have previously shown that the slow Wallerian degeneration mutation, whilst delaying axonal degeneration after optic nerve crush, does not protect retinal ganglion cell (RGC bodies in adult rats. To test the effects of a combination approach protecting both axons and cell bodies we performed combined optic nerve crush and lens injury, which results in both enhanced RGC survival as well as axon regeneration past the lesion site in wildtype animals. Results As previously reported we found that the WldS mutation does not protect RGC bodies after optic nerve crush alone. Surprisingly, we found that WldS transgenic rats did not exhibit the enhanced RGC survival response after combined optic nerve crush and lens injury that was observed in wildtype rats. RGC axon regeneration past the optic nerve lesion site was, however, similar in WldS and wildtypes. Furthermore, activation of retinal glia, previously shown to be associated with enhanced RGC survival and axon regeneration after optic nerve crush and lens injury, was unaffected in WldS transgenic rats. Conclusions RGC axon regeneration is similar between WldS transgenic and wildtype rats, but WldS transgenic rats do not exhibit enhanced RGC survival after combined optic nerve crush and lens injury suggesting that the neuroprotective effects of lens injury on RGC survival may be limited by the WldS protein.

Lorber Barbara

2012-06-01

152

Establishment of an invasive prostate cancer model in transgenic rats by intermittent testosterone administration.  

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We have established a transgenic rat for adenocarcinoma of the prostate (TRAP) model that features uniform adenocarcinoma development in prostatic lobes at high incidence within a short experimental period. However, no invasive carcinomas with reactive stroma characteristics similar to those in man were observed. We therefore have focused on a new model for invasive carcinoma of the prostate using TRAP rats. In experiment 1, male TRAP rats in groups 1 and 2 were treated with orchiectomy at day 0 of the experiment. Rats in groups 1-3 underwent testosterone propionate (TP) implantation from weeks 1 to 4 and from weeks 6 to 16. Rats in groups 1 and 3 were given 3,2'-dimethyl-4-aminobiphenyl (DMAB) after TP implantation. The rats of group 4 served as controls. In experiment 2, the rats were divided into three groups, none of which received DMAB or orchiectomy, treated with TP continuously or with the treatment withdrawn once or twice. In experiment 1, invasive adenocarcinomas with abundant collagenous stroma were found in the dorsolateral and anterior prostate, some of which showed perineural space invasion at week 16. The number of invasive carcinoma foci was most frequent in group 3. In experiment 2, invasive adenocarcinoma development in the lateral prostates was correlated with the number of TP administration/withdrawal cycles. In conclusion, our newly established rat model for invasive adenocarcinoma of the prostate could serve as a useful preclinical model for evaluating the in vivo efficacy of preventive and therapeutic agents targeting of the tumor microenvironment. PMID:24791066

Sato, Shinya; Suzuki, Shugo; Naiki-Ito, Aya; Komiya, Masami; Ne, Long; Kato, Hiroyuki; Sagawa, Hiroyuki; Yamashita, Yoriko; Shirai, Tomoyuki; Takahashi, Satoru

2014-04-01

153

Angiotensin II induced inflammation in the kidney and in the heart of double transgenic rats  

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Full Text Available Abstract Background We are investigating a double transgenic rat (dTGR model, in which rats transgenic for the human angiotensinogen and renin genes are crossed. These rats develop moderately severe hypertension but die of end-organ cardiac and renal damage by week 7. The heart shows necrosis and fibrosis, whereas the kidneys resemble the hemolytic-uremic syndrome vasculopathy. Surface adhesion molecules (ICAM-1 and VCAM-1 are expressed early on the endothelium, while the corresponding ligands are found on circulating leukocytes. Leukocyte infiltration in the vascular wall accompanies PAI-1, MCP-1, iNOS and Tissue Factor expression. Furthermore we show evidence that Ang II causes the upregulation of NF-kB in our model. Methods We started PDTC-treatment on four weeks old dTGR (200 mg/kg sc and age-matched SD rats.. Blood-pressure- and albuminuria- measurements were monitored during the treatement period (four weeks. The seven weeks old animals were killed, hearts and kidneys were isolated and used for immunohistochemical-and electromobility shift assay analsis. Results Chronic treatment with the antioxidant PDTC decreased blood pressure (162 ± 8 vs. 190 ± 7 mm Hg, p = 0.02. Cardiac hypertrophy index was significantly reduced (4.90 ± 0.1 vs. 5.77 ± 0.1 mg/g, p Conclusion Our data show that inhibition of NF-?B by PDTC markedly reduces inflammation, iNOS expression in the dTGR most likely leading to decreased cytotoxicity, and cell proliferation. Thus, NF-?B activation plays an important role in ANG II-induced end-organ damage.

Haller Hermann

2002-01-01

154

Mechanisms of hypertension in transgenic rats expressing the mouse Ren-2 gene.  

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Transgenic (TG) rats carrying the mouse Ren-2 gene (Ren-2d)27 are a newly established monogenetic model in hypertension research. To gain an insight into the mechanisms of this form of hypertension we determined the effects of a 13-day therapy with losartan (10 mg/kg) or lisinopril (20 mg/kg) on the blood pressure (BP) and plasma levels of angiotensin (ANG) peptides of mature female TG hypertensive and Sprague-Dawley (SD) rats. The contribution of endothelium-derived nitric oxide (NO) to the maintenance of their hypertension and the response to therapy was evaluated by systemic injection of either NG-monomethyl-L-arginine (L-NMMA) or endothelin-1. Hypertension in TG rats was associated with decreased plasma ANG I, no differences in plasma ANG II, and plasma ANG-(1-7) near the detectable level. Lisinopril lowered BP more than losartan in both TG hypertensive and normotensive controls. In both strains, the chronic fall in BP produced by lisinopril was accompanied by significant increases in plasma ANG I and ANG-(1-7), while losartan augmented plasma ANG I and ANG II in both strains and plasma ANG-(1-7) in TG rats. Inhibition of NO synthase reversed the fall in BP produced by either lisinopril or losartan in SD controls. In contrast, administration of L-NMMA to TG rats given the same therapy did not. The transient endothelium-mediated relaxing phase of the depressor response to systemic injections of endothelin-1 was attenuated by losartan and lisinopril in TG rats. These studies indicate that hypertension in female TG rats is mediated by the RAS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8184972

Moriguchi, A; Brosnihan, K B; Kumagai, H; Ganten, D; Ferrario, C M

1994-04-01

155

A progressive dopaminergic phenotype associated with neurotoxic conversion of ?-synuclein in BAC-transgenic rats  

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Conversion of soluble ?-synuclein into insoluble and fibrillar inclusions is a hallmark of Parkinson's disease and other synucleinopathies. Accumulating evidence points towards a relationship between its generation at nerve terminals and structural synaptic pathology. Little is known about the pathogenic impact of ?-synuclein conversion and deposition at nigrostriatal dopaminergic synapses in transgenic mice, mainly owing to expression limitations of the ?-synuclein construct. Here, we explore whether both the rat as a model and expression of the bacterial artificial chromosome construct consisting of human full-length wild-type ?-synuclein could exert dopaminergic neuropathological effects. We found that the human promoter induced a pan-neuronal expression, matching the rodent ?-synuclein expression pattern, however, with prominent C-terminally truncated fragments. Ageing promoted conversion of both full-length and C-terminally truncated ?-synuclein species into insolube and proteinase K-resistant fibres, with strongest accumulation in the striatum, resembling biochemical changes seen in human Parkinson's disease. Transgenic rats develop early changes in novelty-seeking, avoidance and smell before the progressive motor deficit. Importantly, the observed pathological changes were associated with severe loss of the dopaminergic integrity, thus resembling more closely the human pathology.

Nuber, Silke; Harmuth, Florian

2013-01-01

156

Inducible and reversible gene silencing by stable integration of an shRNA-encoding lentivirus in transgenic rats  

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Currently, tools to generate loss-of-function mutations in rats are limited. Therefore, we have developed a lentiviral single-vector system for the temporal control of ubiquitous shRNA expression. Here, we report transgenic rats carrying an insulin receptor-specific shRNA transcribed from a regulatable promoter and identified by concomitant EGFP expression. In the absence of the inducer doxycycline (Dox), we observed no siRNA expression. However, Dox treatment at very low concentrations led t...

Herold, Marco J.; Den Brandt, Jens; Seibler, Jost; Reichardt, Holger M.

2008-01-01

157

Transgenic analysis of the response of the rat calbindin-D 9k gene to vitamin D.  

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The promoter of the calbindin-D 9k (CaBP9k) gene, previously analyzed in transgenic mice, contains all of the information necessary for expression of a transgene similar to the endogenous gene and also for an appropriate response to vitamin D. In the present study we first investigated the role of a putative vitamin D-responsive element (9k/VDRE), located at nucleotides -489 to -445 on the rat CaBP9k promoter gene, using transgenic mice. As expected, the pattern of transgene expression in mice carrying this putative VDRE mutated in its whole promoter context was similar to that in mice bearing the wild-type sequence. These transgenic mice also responded to 1,25-dihydroxyvitamin D3 in the same way as those bearing the wild-type transgene and as those carrying a transgene with a large deletion (from -2894 to -117) eliminating the putative 9k/VDRE. Thus, the putative 9k/VDRE is not required for the control of rat CaBP9k gene expression by vitamin D in vivo. We also found that responsiveness to 1,25-dihydroxyvitamin D3 depends on the site at which the transgene is integrated into the host genome, in a tissue-specific manner. These data together with the fact that vitamin D-responsive sequences are present in a two-module region (from -3731 to -2894 and/or -117 to +365) and that this region does not contain any classical VDRE show that the CaBP9k gene is submitted to a non-conventional control by vitamin D. PMID:10875229

Colnot, S; Ovejero, C; Romagnolo, B; Porteu, A; Lacourte, P; Thomasset, M; Perret, C

2000-07-01

158

Modified impact of emotion on temporal discrimination in a transgenic rat model of Huntington disease  

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Full Text Available Huntington’s disease (HD is characterized by triad of motor, cognitive and emotional symptoms along with neuropathology in fronto-striatal circuit and limbic system including amygdala. Emotional alterations, which have a negative impact on patient well-being, represent some of the earliest symptoms of HD and might be related to the onset of the neurodegenerative process. In the transgenic rat model (tgHD rats, evidence suggest emotional alterations at the symptomatic stage along with neuropathology of the central nucleus of amygdala (CE. Studies in humans and animals demonstrate that emotion can modulate time perception. The impact of emotion on time perception has never been tested in HD, nor is it known if that impact could be part of the presymptomatic emotional phenotype of the pathology. The aim of this paper was to characterize the effect of emotion on temporal discrimination in presymptomatic tgHD animals. In the first experiment, we characterized the acute effect of an emotion (fear conditioned stimulus on temporal discrimination using a bisection procedure, and tested its dependency upon an intact central amygdala. The second experiment was aimed at comparing presymptomatic homozygous transgenic animals at 7-months of age and their wild-type littermates (WT in their performance on the modulation of temporal discrimination by emotion. Our principal findings show that (1 a fear cue produces a short-lived decrease of temporal precision after its termination, and (2 animals with medial CE lesion and presymptomatic tgHD animals demonstrate an alteration of this emotion-evoked temporal distortion. The results contribute to our knowledge about the presymptomatic phenotype of this HD rat model, showing susceptibility to emotion that may be related to dysfunction of the central nucleus of amygdala.

AlexisFaure

2013-09-01

159

Changes in endovascular trophoblast invasion and spiral artery remodelling at term in a transgenic preeclamptic rat model.  

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As a follow-up to our previous study which revealed a surprisingly deeper endovascular trophoblast (ET) invasion on day 18 in a transgenic preeclamptic (PE) rat model (hAngiotensinogen female symbol x hRenin male symbol) compared to non-PE controls, we examined further changes in ET invasion and associated spiral artery (SA) remodelling at term (day 21). PE transgenic rats and non-PE reversely mated (RM) transgenic rats were compared to normal SD rats (C). Sections were stained to visualize trophoblast, fibrinoid, vascular smooth muscle (VSM) and endothelium. SA were evaluated in three depth levels in the mesometrial triangle (MT) using the KS-400 image analysis system. In separate transgenic rats, Doppler ultrasound was performed in uterine arteries, and the resistance indices (RI) were calculated. Although for the whole MT differences in ET invasion were no longer significant between the PE and C, indicating a partial catching up in C rats, there was still significantly more ET in the deepest level in the PE group as compared to the C and RM groups. At the same time the SA walls in PE rats contained significantly more fibrinoid (versus RM and C) and VSM (versus C). In all SA cross-sections, re-endothelialisation was prominent, but significantly different between PE and C group. The Doppler results showed a significantly lower RI in the arcuate uterine artery of the PE group compared to the C group. There was no evidence of elimination of deeply invaded ET at term, previously considered as a possible mechanism for restriction of vascular remodelling in human PE. The differences in vascular remodelling, previously described on day 18 by histology and Doppler data, were maintained on day 21, but there was extensive endothelial repair in the three groups. Atherosis-like lesions were observed in the three groups, most frequently in the RM group, but were never associated with placental infarcts. PMID:20144482

Geusens, N; Hering, L; Verlohren, S; Luyten, C; Drijkoningen, K; Taube, M; Vercruysse, L; Hanssens, M; Dechend, R; Pijnenborg, R

2010-04-01

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Transgenic overexpression of the sarcoplasmic reticulum Ca2+ATPase improves reticular Ca2+ handling in normal and diabetic rat hearts.  

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Slowed relaxation in diabetic cardiomyopathy (CM) is partially related to diminished expression of the sarcoplasmic reticulum (SR) Ca2+-ATPase SERCA2a. To evaluate the impact of SERCA2a overexpression on SR Ca2+ handling in diabetic CM, we 1) generated transgenic rats harboring a human cytomegalovirus enhancer/chicken beta-actin promotor-controlled rat SERCA2 transgene (SERCA2-TGR), 2) characterized their SR phenotype, and 3) examined whether transgene expression may rescue SR Ca2+ transport in streptozotocin-induced diabetes. The transgene was expressed in all heart chambers. Compared to wild-type (WT) rats, a heterozygous line exhibited increased SERCA2 mRNA (1.5-fold), SERCA2 protein (+26%) and SR Ca2+ uptake (+37%). Phospholamban expression was not altered. In SERCA2-TGR, contraction amplitude (+48%) and rates of contraction (+34%) and relaxation (+35%) of isolated papillary muscles (PM) were increased (P2+ uptake and SERCA2 protein of SERCA2-TGR were 1.3-fold higher (P2+ uptake, accelerates relaxation and compensates, in part, for depressed Ca2+ uptake in diabetic CM. Therefore, SERCA2 expression might constitute an important therapeutic target to rescue cardiac SR Ca2+ handling in diabetes. PMID:12206992

Vetter, Roland; Rehfeld, Uwe; Reissfelder, Christoph; Weiss, Wolfgang; Wagner, Kay-Dietrich; Günther, Joachim; Hammes, Annette; Tschöpe, Carsten; Dillmann, Wolfgang; Paul, Martin

2002-10-01

 
 
 
 
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Autonomic cardiac control in animal models of cardiovascular diseases II. Variability analysis in transgenic rats with alpha-tropomyosin mutations Asp175Asn and Glu180Gly  

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Animal models of cardiovascular diseases allow to investigate relevant pathogenetic mechanisms in detail. In the present study, the mutations Asp175Asn and Glu180Gly in alpha-tropomyosin (TPM1), known cause familiar hypertrophic cardiomyopathy (FHC) were studied for changes in hemodynamic parameters and spontaneous baroreflex regulation in transgenic rats in comparison to transgenic and non-transgenic controls by telemetry. Heart rate variability (HRV) and blood pressure variability (BPV) wer...

Wernicke, D.; Wessel, N.; Malberg, H.; Plehm, R.; Bauernschmitt, R.; Thierfelder, L.

2007-01-01

162

Leflunomide derivate FK 778 in accelerated renal injury in transgenic rat.  

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Renal ischaemia/reperfusion (I/R) injury and hypertension represent major alloantigen-independent risk factors contributing to the development of chronic allograft nephropathy. In a model of accelerated major histocompatibility complex-independent renal injury, we evaluated the effect of leflunomide derivate - FK778 - on the progression of accelerated nephropathy. Thirty-six uninephrectomized hypertensive transgenic (m-REN-2)-27 rats received a clip on renal pedicle for 45 minutes. Animals were treated with FK778 3 mg/kg/day (I/R 3 mg, N = 12), 10 mg/kg/day (I/R 10 mg, N = 12) or placebo (N = 12) via gavage for 16 weeks. Eighteen animals were sham-operated and treated with FK778 3 mg/kg/day (sham 3 mg, N = 6), 10 mg/kg/day (sham 10 mg, N = 6) or were untreated (sham, N = 6). Proteinuria and blood pressure were evaluated throughout and the kidneys were harvested for morphological and immunohistochemical analysis at the end of the experiment. At week 16, rats with I/R injury and FK778 treatment had lower proteinuria compared with placebo-treated rats (I/R 3 mg: 48.42 +/- 26.16, I/R 10 mg 27.28 +/- 21.86 vs. Placebo: 70.13 +/- 50.19 mg/day, P FK778-treated sham groups (Sham 3 mg: 24.23 +/- 10.89; Sham 10 mg: 17.37 +/- 4.13; Sham: 14.23 +/- 1.18) There was no difference in glomerulosclerosis and interstitial fibrosis among the treated groups. In the untreated animals the rate of interstitial fibrosis decline reached statistical significance (Placebo vs. Sham: 1.125 +/- 0.641 % vs. 0.250 +/- 0.500 %, P FK778-treated rats displayed amelioration of some changes induced by the I/R injury. Our observation also suggests potential nephrotoxicity of FK778. PMID:20492759

Bloudícková, S; Rajnoch, J; Lodererová, A; Honsová, E; Viklický, O

2010-01-01

163

Meloxicam Blocks Neuroinflammation, but Not Depressive-Like Behaviors, in HIV-1 Transgenic Female Rats  

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Adolescents living with human immunodeficiency virus (HIV) comprise approximately 12% of the HIV-positive population worldwide. HIV-positive adolescents experience a higher rate of clinical depression, a greater risk of sexual and drug abuse behaviors, and a decreased adherence to highly active antiretroviral therapies (HAART). Using adolescent HIV-1 transgenic rats (HIV-1 tg) that display related immune response alterations and pathologies, this study tested the hypothesis that developmental expression of HIV-1-related proteins induces a depressive-like phenotype that parallels a decrease in hippocampal cell proliferation and an increase in pro-inflammatory cytokine expression in the hippocampus. Consistent with this hypothesis, adolescent HIV-1 tg rats demonstrated a depressive-like behavioral phenotype, had decreased levels of cell proliferation, and exhibited elevated expression of monocyte chemotactic protein-1 (Mcp-1) in the hippocampus relative to controls. Subsequently, we tested the ability of meloxicam, a selective COX-2 inhibitor, to attenuate behavioral deficits via inflammatory mechanisms. Daily meloxicam treatments did not alter the behavioral profile despite effectively reducing hippocampal inflammatory gene expression. Together, these data support a biological basis for the co-morbid manifestation of depression in HIV-positive patients as early as in adolescence and suggest that modifications in behavior manifest independent of inflammatory activity in the hippocampus. PMID:25271421

Nemeth, Christina L.; Glasper, Erica R.; Harrell, Constance S.; Malviya, Sanjana A.; Otis, Jeffrey S.; Neigh, Gretchen N.

2014-01-01

164

Food-anticipatory activity and liver per1-luc activity in diabetic transgenic rats  

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The mammalian Per1 gene is an important component of the core cellular clock mechanism responsible for circadian rhythms. The rodent liver and other tissues rhythmically express Per1 in vitro but typically damp out within a few cycles. In the liver, the peak of this rhythm occurs in the late subjective night in an ad lib-fed rat, but will show a large phase advance in response to restricted availability of food during the day. The relationship between this shift in the liver clock and food-anticipatory activity (FAA), the circadian behavior entrained by daily feeding, is currently unknown. Insulin is released during feeding in mammals and could serve as an entraining signal to the liver. To test the role of insulin in the shift in liver Per1 expression and the generation of FAA, per-luciferase transgenic rats were made diabetic with a single injection of streptozotocine. Following 1 week of restricted feeding and locomotor activity monitoring, liver was collected for per-luc recording. In two separate experiments, FAA emerged and liver Per1 phase-shifted in response to daytime 8-h food restriction. The results rule out insulin as a necessary component of this system.

Davidson, Alec J.; Stokkan, Karl-Arne; Yamazaki, Shin; Menaker, Michael

2002-01-01

165

Transgenic rats overexpressing the human MrgX3 gene show cataracts and an abnormal skin phenotype  

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The human MrgX3 gene, belonging to the mrgs/SNSRs (mass related genes/sensory neuron specific receptors) family, was overexpressed in transgenic rats using the actin promoter. Two animal lines showed cataracts with liquification/degeneration and swelling of the lens fiber cells. The transient epidermal desquamation was observed in line with higher gene expression. Histopathology of the transgenic rats showed acanthosis and focal parakeratosis. In the epidermis, there was an increase in cellular keratin 14, keratin 10, and loricrin, as well as PGP 9.5 in innervating nerve fibers. These phenotypes accompanied an increase in the number of proliferating cells. These results suggest that overexpression of the human MrgX3 gene causes a disturbance of the normal cell-differentiation process

166

Partial redirection of transgenic human growth hormone secretion from rat salivary glands.  

Science.gov (United States)

Regulated secretory pathway proteins, when delivered as transgenes to salivary glands, are secreted predominantly into saliva. This is not useful for those proteins whose therapeutic function is required systemically, for example, human growth hormone (hGH). One strategy to improve the efficiency of hGH secretion into the bloodstream involves manipulation of existing sorting signals. The C terminus of hGH is highly conserved and contains a domain similar to the regulated pathway sorting domain of pro-opiomelanocortin (POMC). We hypothesized that, similar to POMC, mutation of this domain would divert hGH secretion from the regulated to the constitutive pathway, which in salivary glands leads to the bloodstream. Several mutations were made in the C terminus of the hGH cDNA and tested in vitro. One biologically active mutant containing E174A and E186A substitutions, and with an included C-terminal extension, was studied in greater detail. Compared with wild-type hGH, we found that this mutant hGH accumulated in the Golgi/trans-Golgi network and showed increased basal secretion in AtT20 cells, a model endocrine cell line. Importantly, in vivo, the mutant hGH displayed a relative increase in the proportion of constitutive pathway secretion seen from rat salivary glands, with a significantly lower saliva-versus-serum secretion ratio (p=0.03). Although this mutant is unlikely to be therapeutically beneficial, these results suggest that the final destination of a transgenic secretory protein may be controlled by reengineering its sorting determinants. PMID:15916482

Wang, Jianghua; Cawley, Niamh X; Voutetakis, Antonis; Rodriguez, Yazmin M; Goldsmith, Corinne M; Nieman, Lynnette K; Hoque, A T M Shamsul; Frank, Stuart J; Snell, Chris R; Loh, Y Peng; Baum, Bruce J

2005-05-01

167

Vitamin A deficiency and behavioral and motor deficits in the human immunodeficiency virus type 1 transgenic rat.  

Science.gov (United States)

The human immunodeficiency virus type 1 (HIV-1) transgenic (Tg) rat model incorporates a noninfectious viral genome that is under similar regulatory control mechanisms in vivo as those that exist with natural infection in humans. Vitamin A (VA) deficiency in humans has been associated with progressive systemic HIV disease and with impaired cognition in rodent models. The effects on of VA deficiency on the development of behavioral abnormalities with HIV infection have not been previously described. In these studies, wild-type (Wt) and Tg rats maintained on either a normal (VA+) or a VA-deficient (VA-) diet were examined for activity in an open field (horizontal activity, total distance, vertical activity, and rearing) and on rotarod testing. On both open field and rotarod testing, the Tg rats performed worse than the Wt rats, with the most severe deficits noted in the TgVA- animals. Analysis of the specific effects of the presence of the HIV transgene and the diet on the performance on the open field tests showed a dominant effect from the transgene on all of the tests, with an effect from the diet on only the number of rearings. On rotarod testing, effects form both the diet and the transgene were observed at lower speeds, at the highest speeds, and on the accelerating rotarod. These studies therefore demonstrate that behavioral and motor abnormalities can be detected in this model and are likely due to similar mechanisms by which humans infected with HIV might develop cognitive-motor impairment in association with VA deficiency. PMID:19995129

June, Harry L; Tzeng Yang, Andrew Rong Song; Bryant, Joseph L; Jones, Odell; Royal, Walter

2009-09-01

168

Adolescent HIV-1 Transgenic Rats: Evidence for Dopaminergic Alterations in Behavior and Neurochemistry Revealed by Methamphetamine Challenge  

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Since the introduction of combination antiretroviral therapy (cART) in the mid-90s, the most severe forms of HIV-1-associated neurocognitive disorders (HAND) have diminished. However, milder forms of HAND remain prevalent. Basic and clinical studies implicate alterations in the dopaminergic (DAergic) system in HIV-1 infection. We used the Fischer 344 HIV-1 transgenic (HIV-1 Tg) rat, which expresses 7 of the 9 HIV-1 genes, to examine potential DAergic alterations. Animals were studied beginnin...

Moran, Landhing M.; Aksenov, Michael Y.; Booze, Rosemarie M.; Webb, Katy M.; Mactutus, Charles F.

2012-01-01

169

Optogenetic patterning of whisker-barrel cortical system in transgenic rat expressing channelrhodopsin-2.  

Science.gov (United States)

The rodent whisker-barrel system has been an ideal model for studying somatosensory representations in the cortex. However, it remains a challenge to experimentally stimulate whiskers with a given pattern under spatiotemporal precision. Recently the optogenetic manipulation of neuronal activity has made possible the analysis of the neuronal network with precise spatiotemporal resolution. Here we identified the selective expression of channelrhodopsin-2 (ChR2), an algal light-driven cation channel, in the large mechanoreceptive neurons in the trigeminal ganglion (TG) as well as their peripheral nerve endings innervating the whisker follicles of a transgenic rat. The spatiotemporal pattern of whisker irradiation thus produced a barrel-cortical response with a specific spatiotemporal pattern as evidenced by electrophysiological and functional MRI (fMRI) studies. Our methods of generating an optogenetic tactile pattern (OTP) can be expected to facilitate studies on how the spatiotemporal pattern of touch is represented in the somatosensory cortex, as Hubel and Wiesel did in the visual cortex. PMID:24695456

Honjoh, Tatsuya; Ji, Zhi-Gang; Yokoyama, Yukinobu; Sumiyoshi, Akira; Shibuya, Yuma; Matsuzaka, Yoshiya; Kawashima, Ryuta; Mushiake, Hajime; Ishizuka, Toru; Yawo, Hiromu

2014-01-01

170

Sustained and promoter dependent bone morphogenetic protein expression by rat mesenchymal stem cells after BMP-2 transgene electrotransfer  

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Full Text Available Transplantation of mesenchymal stem cells (MSCs with electrotransferred bone morphogenetic protein-2 (BMP-2 transgene is an attractive therapeutic modality for the treatment of large bone defects: it provides both stem cells with the ability to form bone and an effective bone inducer while avoiding viral gene transfer. The objective of the present study was to determine the influence of the promoter driving the human BMP-2 gene on the level and duration of BMP-2 expression after transgene electrotransfer into rat MSCs. Cytomegalovirus, elongation factor-1?, glyceraldehyde 3-phosphate dehydrogenase, and beta-actin promoters resulted in a BMP-2 secretion rate increase of 11-, 78-, 66- and 36-fold over respective controls, respectively. In contrast, the osteocalcin promoter had predictable weak activity in undifferentiated MSCs but induced the strongest BMP-2 secretion rates in osteoblastically-differentiated MSCs. Regardless of the promoter driving the transgene, a plateau of maximal BMP-2 secretion persisted for at least 21 d after the hBMP-2 gene electrotransfer. The present study demonstrates the feasibility of gene electrotransfer for efficient BMP-2 transgene delivery into MSCs and for a three-week sustained BMP-2 expression. It also provides the first in vitro evidence for a safe alternative to viral methods that permit efficient BMP-2 gene delivery and expression in MSCs but raise safety concerns that are critical when considering clinical applications.

E Ferreira

2012-07-01

171

Distinct Transcriptional Mechanisms Direct Expression of the Rat Dmrt1 Promoter in Sertoli Cells and Germ Cells of Transgenic Mice1  

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DMRT1 is a transcription factor expressed only in Sertoli cells and undifferentiated spermatogonia of the postnatal testis, where it is required for proper cellular differentiation and fertility. To elucidate the transcriptional regulatory regions that provide DMRT1's cell-specific expression, transgenic mice containing a LacZ reporter gene driven by variable amounts of rat Dmrt1 5? flanking sequence, 9 kb and smaller, were evaluated. Examination of transgene expression by RT-PCR indicated ...

Lei, Ning; Karpova, Tatiana; Hornbaker, Kaori I.; Rice, Daren A.; Heckert, Leslie L.

2009-01-01

172

Prostate and mammary adenocarcinoma in transgenic mice carrying a rat C3(1) simian virus 40 large tumor antigen fusion gene.  

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A transgenic mouse model for prostate and mammary cancer has been developed in mice containing a recombinant gene expressing the simian virus 40 early-region transforming sequences under the regulatory control of the rat prostatic steroid binding protein [C3(1)] gene. Male transgenic mice develop prostatic hyperplasia in early life that progresses to adenoma or adenocarcinoma in most animals surviving to longer than 7 months of age. Prostate cancer metastases to lung have been observed. Femal...

Maroulakou, I. G.; Anver, M.; Garrett, L.; Green, J. E.

1994-01-01

173

Visual properties of transgenic rats harboring the channelrhodopsin-2 gene regulated by the thy-1.2 promoter.  

Science.gov (United States)

Channelrhodopsin-2 (ChR2), one of the archea-type rhodopsins from green algae, is a potentially useful optogenetic tool for restoring vision in patients with photoreceptor degeneration, such as retinitis pigmentosa. If the ChR2 gene is transferred to retinal ganglion cells (RGCs), which send visual information to the brain, the RGCs may be repurposed to act as photoreceptors. In this study, by using a transgenic rat expressing ChR2 specifically in the RGCs under the regulation of a Thy-1.2 promoter, we tested the possibility that direct photoactivation of RGCs could restore effective vision. Although the contrast sensitivities of the optomotor responses of transgenic rats were similar to those observed in the wild-type rats, they were enhanced for visual stimuli of low-spatial frequency after the degeneration of native photoreceptors. This result suggests that the visual signals derived from the ChR2-expressing RGCs were reinterpreted by the brain to form behavior-related vision. PMID:19893752

Tomita, Hiroshi; Sugano, Eriko; Fukazawa, Yugo; Isago, Hitomi; Sugiyama, Yuka; Hiroi, Teru; Ishizuka, Toru; Mushiake, Hajime; Kato, Megumi; Hirabayashi, Masumi; Shigemoto, Ryuichi; Yawo, Hiromu; Tamai, Makoto

2009-01-01

174

Dynamics of testicular germ cell apoptosis in normal mice and transgenic mice overexpressing rat androgen-binding protein  

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Abstract The number and type of testicular germ cells undergoing apoptosis in different age groups of mice (from 7 to 360 days of age) was determined and compared in age-matched wild type (WT) control and in a transgenic (TG) mice homozygous to rat androgen binding protein (ABP) using flow cytometry. Flow cytometric quantification revealed that the total number of germ cells undergoing apoptosis did not differ significantly in WT and TG mice up to Day 14. From Day 21 to Day 60, the ...

Petrusz Peter; Grossman Gail; Antony, Jeyaraj D.

2003-01-01

175

Highly visible expression of an oxytocin-monomeric red fluorescent protein 1 fusion gene in the hypothalamus and posterior pituitary of transgenic rats.  

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We have generated rats bearing an oxytocin (OXT)-monomeric red fluorescent protein 1 (mRFP1) fusion transgene. The mRFP1 fluorescence was highly visible in ventral part of the supraoptic nucleus (SON) and the posterior pituitary in a whole mount. mRFP1 fluorescence in hypothalamic sections was also observed in the SON, the paraventricular nucleus (PVN), and the internal layer of the median eminence. Salt loading for 5 d caused a marked increase in mRFP1 fluorescence in the SON, the PVN, the median eminence, and the posterior pituitary. In situ hybridization histochemistry revealed that the expression of the mRNA encoding the OXT-mRFP1 fusion gene was observed in the SON and the PVN of euhydrated rats and increased dramatically after chronic salt loading. The expression of the endogenous OXT and the arginine vasopressin (AVP) genes were significantly increased in the SON and the PVN after chronic salt loading in both nontransgenic and transgenic rats. These responses were not different between male and female rats. Compared with nontransgenic rats, euhydrated and salt-loaded male and female transgenic rats showed no significant differences in plasma osmolality, sodium concentration, OXT, and AVP levels. Finally, we succeeded in generating a double-transgenic rat that expresses both the OXT-mRFP1 fusion gene and the AVP-enhanced green fluorescent protein fusion gene. Our new transgenic rats are valuable new tools to study the physiology of the hypothalamo-neurohypophysial system. PMID:21540286

Katoh, Akiko; Fujihara, Hiroaki; Ohbuchi, Toyoaki; Onaka, Tatsushi; Hashimoto, Takashi; Kawata, Mitsuhiro; Suzuki, Hideaki; Ueta, Yoichi

2011-07-01

176

Development and characterization of a new inbred transgenic rat strain expressing DsRed monomeric fluorescent protein.  

Science.gov (United States)

The inbred rat is a suitable model for studying human disease and because of its larger size is more amenable to complex surgical manipulation than the mouse. While the rodent fulfills many of the criteria for transplantation research, an important requirement is the ability to mark and track donors cells and assess organ viability. However, tracking ability is limited by the availability of transgenic (Tg) rats that express suitable luminescent or fluorescent proteins. Red fluorescent protein cloned from Discosoma coral (DsRed) has several advantages over other fluorescent proteins, including in vivo detection in the whole animal and ex vivo visualization in organs as there is no interference with autofluorescence. We generated and characterized a novel inbred Tg Lewis rat strain expressing DsRed monomeric (DsRed mono) fluorescent protein under the control of a ubiquitously expressed ROSA26 promoter. DsRed mono Tg rats ubiquitously expressed the marker gene as detected by RT-PCR but the protein was expressed at varying levels in different organs. Conventional skin grafting experiments showed acceptance of DsRed monomeric Tg rat skin on wild-type rats for more than 30 days. Cardiac transplantation of DsRed monomeric Tg rat hearts into wild-type recipients further showed graft acceptance and long-term organ viability (>6 months). The DsRed monomeric Tg rat provides marked cells and/or organs that can be followed for long periods without immune rejection and therefore is a suitable model to investigate cell tracking and organ transplantation. PMID:25011565

Montanari, Sonia; Wang, Xing-Hua; Yannarelli, Gustavo; Dayan, Victor; Berger, Thorsten; Zocche, Larissa; Kobayashi, Eiji; Viswanathan, Sowmya; Keating, Armand

2014-10-01

177

Elevated suppressor of cytokine signaling-1 (SOCS-1): a mechanism for dysregulated osteoclastogenesis in HIV transgenic rats.  

Science.gov (United States)

Accelerated bone loss leading to osteopenia, osteoporosis, and bone fracture is a major health problem that is increasingly common in human immunodeficiency virus (HIV)-infected patients. The underlying pathogenesis is unclear but occurs in both treatment naïve and individuals receiving antiretroviral therapies. We developed an HIV-1 transgenic rat that exhibits many key features of HIV disease including HIV-1-induced changes in bone mineral density (BMD). A key determinant in the rate of bone loss is the differentiation of osteoclasts, the cells responsible for bone resorption. We found HIV-1 transgenic osteoclast precursors (OCP) express higher levels of suppressor of cytokine signaling-1 (SOCS-1) and TNF receptor-associated factor 6 (TRAF6) and are resistant to interferon-gamma (IFN-?) mediated suppression of osteoclast differentiation. Our data suggest that dysregulated SOCS-1 expression by HIV-1 transgenic OCP promotes osteoclastogenesis leading to the accelerated bone loss observed in this animal model. We propose that elevated SOCS-1 expression in OCP antagonizes the inhibitory effects of IFN-? and enhances receptor activator of NF-kB ligand (RANKL) signaling that drives osteoclast differentiation and activation. Understanding the molecular mechanisms of HIV-associated BMD changes has the potential to detect and treat bone metabolism disturbances early and improve the quality of life in patients. PMID:24376119

Lafferty, Mark K; Fantry, Lori; Bryant, Joseph; Jones, Odell; Hammoud, Dima; Weitzmann, M Neale; Lewis, George K; Garzino-Demo, Alfredo; Reid, William

2014-06-01

178

Characterization of dsRed2-positive cells in the doublecortin-dsRed2 transgenic adult rat retina.  

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Doublecortin (DCX) is predominantly expressed in neuronal precursor cells and young immature neurons of the developing and adult brain, where it is involved in neuronal differentiation, migration and plasticity. Moreover, its expression pattern reflects neurogenesis, and transgenic DCX promoter-driven reporter models have been previously used to investigate adult neurogenesis. In this study, we characterize dsRed2 reporter protein-expressing cells in the adult retina of the transgenic DCX promoter-dsRed2 rat model, with the aim to identify cells with putative neurogenic activity. Additionally, we confirmed the expression of the dsRed2 protein in DCX-expressing cells in the adult hippocampal dentate gyrus. Adult DCX-dsRed2 rat retinas were analyzed by immunohistochemistry for expression of DCX, NF200, Brn3a, Sox2, NeuN, calbindin, calretinin, PKC-a, Otx2, ChAT, PSA-NCAM and the glial markers GFAP and CRALBP, followed by confocal laser-scanning microscopy. In addition, brain sections of transgenic rats were analyzed for dsRed2 expression and co-localization with DCX, NeuN, GFAP and Sox2 in the cortex and dentate gyrus. Endogenous DCX expression in the adult retina was confined to horizontal cells, and these cells co-expressed the DCX promoter-driven dsRed2 reporter protein. In addition, we encountered dsRed2 expression in various other cell types in the retina: retinal ganglion cells (RGCs), a subpopulation of amacrine cells, a minority of bipolar cells and in perivascular cells. Since also RGCs expressed dsRed2, the DCX-dsRed2 rat model might offer a useful tool to study RGCs in vivo under various conditions. Müller glial cells, which have previously been identified as cells with stem cell features and with neurogenic potential, did express neither endogenous DCX nor the dsRed2 reporter. However, and surprisingly, we identified a perivascular glial cell type expressing the dsRed2 reporter, enmeshed with the glia/stem cell marker GFAP and colocalizing with the neural stem cell marker Sox2. These findings suggest the so far undiscovered existence of perivascular associated cell with neural stem cell-like properties in the adult retina. PMID:25138677

Trost, A; Schroedl, F; Marschallinger, J; Rivera, F J; Bogner, B; Runge, C; Couillard-Despres, S; Aigner, L; Reitsamer, H A

2014-12-01

179

Light-evoked somatosensory perception of transgenic rats that express channelrhodopsin-2 in dorsal root ganglion cells.  

Science.gov (United States)

In vertebrate somatosensory systems, each mode of touch-pressure, temperature or pain is sensed by sensory endings of different dorsal root ganglion (DRG) neurons, which conducted to the specific cortical loci as nerve impulses. Therefore, direct electrical stimulation of the peripheral nerve endings causes an erroneous sensation to be conducted by the nerve. We have recently generated several transgenic lines of rat in which channelrhodopsin-2 (ChR2) transgene is driven by the Thy-1.2 promoter. In one of them, W-TChR2V4, some neurons were endowed with photosensitivity by the introduction of the ChR2 gene, coding an algal photoreceptor molecule. The DRG neurons expressing ChR2 were immunohistochemically identified using specific antibodies to the markers of mechanoreceptive or nociceptive neurons. Their peripheral nerve endings in the plantar skin as well as the central endings in the spinal cord were also examined. We identified that ChR2 is expressed in a certain population of large neurons in the DRG of W-TChR2V4. On the basis of their morphology and molecular markers, these neurons were classified as mechanoreceptive but not nociceptive. ChR2 was also distributed in their peripheral sensory nerve endings, some of which were closely associated with CK20-positive cells to form Merkel cell-neurite complexes or with S-100-positive cells to form structures like Meissner's corpuscles. These nerve endings are thus suggested to be involved in the sensing of touch. Each W-TChR2V4 rat showed a sensory-evoked behavior in response to blue LED flashes on the plantar skin. It is thus suggested that each rat acquired an unusual sensory modality of sensing blue light through the skin as touch-pressure. This light-evoked somatosensory perception should facilitate study of how the complex tactile sense emerges in the brain. PMID:22412908

Ji, Zhi-Gang; Ito, Shin; Honjoh, Tatsuya; Ohta, Hiroyuki; Ishizuka, Toru; Fukazawa, Yugo; Yawo, Hiromu

2012-01-01

180

Differences in acid-induced currents between oxytocin-mRFP1 and vasopressin-eGFP neurons isolated from the supraoptic and paraventricular nuclei of transgenic rats.  

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The hypothalamic supraoptic nucleus (SON) and paraventricular nucleus (PVN) consists of two types of magnocellular neurosecretory cells, oxytocin (OXT) and arginine vasopressin (AVP). We generated and characterized rats that express an OXT-monomeric red fluorescent protein 1 (mRFP1) and an AVP-enhanced green fluorescent protein (eGFP) fusion transgene. These transgenic rats enable the visualization of OXT or AVP neurons. Taking advantage of this, we examined the differences between OXT-mRFP1 neurons and AVP-eGFP neurons in response to acid. Acid-sensing ion channels (ASICs) are neuronal voltage-insensitive cationic channels that are activated by extracellular acidification. Although functional ASICs have been identified in AVP neurons, differences in acid-induced currents between OXT and AVP neurons in SON have not been reported. In the present study, we used the whole-cell patch-clamp technique to investigate differences between OXT-mRFP1 neurons and AVP-eGFP neurons reaction to acid in SON and PVN. In voltage clamp mode, lowering extracellular pH evoked inward currents in both OXT-mRFP1 neurons and AVP-eGFP neurons. In our findings, the acid-induced currents in the OXT-mRFP1 neurons were significantly smaller than those in the AVP-eGFP neurons. These acid-induced currents were inhibited by amiloride, a known blocker of ASICs. Further, to compare the response to acid between OXT-mRFP1 and AVP-eGFP neurons in the same transgenic rat, we used a double transgenic rat by mating an OXT-mRFP1 transgenic rat with an AVP-eGFP transgenic rat. The acid-induced currents of OXT-mRFP1 neurons were significantly smaller than those of AVP-eGFP neurons from the double transgenic rats. These currents were almost completely inhibited by amiloride. The difference of acid-sensitivity between OXT and AVP neurons might contribute to maintaining systematic order in hypothalamic function. PMID:25220704

Ohkubo, Jun-ichi; Ohbuchi, Toyoaki; Yoshimura, Mitsuhiro; Maruyama, Takashi; Hashimoto, Hirofumi; Matsuura, Takanori; Suzuki, Hideaki; Ueta, Yoichi

2014-11-01

 
 
 
 
181

Behavioral, neurochemical, and pathologic alterations in bacterial artificial chromosome transgenic G2019S leucine-rich repeated kinase 2 rats.  

Science.gov (United States)

Mutations in leucine-rich repeated kinase 2 (LRRK2) cause autosomal dominant late-onset Parkinson's disease (PD), and the G2019S mutation in the kinase domain of LRRK2 is the most common genetic cause of familial PD. Enhanced kinase activity of G2019S LRRK2 is a suspected mechanism for carriers to develop PD but pathophysiological function of G2019S LRRK2 is not clear. The objective of the present study was to characterize a bacterial artificial chromosome rat expressing human G2019S LRRK2. Immunoblotting analysis showed that G2019S LRRK2 expression was approximately 5-8 times higher than wild-type rat LRRK2. At ages of 4, 8, and 12 months, our characterization showed that expression of G2019S LRRK2 induced oxidative stress in striatum and substantia nigra, increased inducible nitric oxide synthase expression in nigral dopamine neurons, and abnormal morphology of nigral dopaminergic neurons in transgenic rats compared with wild-type, without inducing overt neurodegeneration in nigrostriatal dopaminergic neurons. Thus, we conclude that although this model does not reproduce the key features of end-stage PD, important preclinical features of the disease are evident, which may be useful in studying the earliest stages of PD and for gene-environment interaction studies. PMID:25174649

Lee, Jang-Won; Tapias, Victor; Di Maio, Roberto; Greenamyre, J Timothy; Cannon, Jason R

2015-01-01

182

Role of HIV-1 Infection in Addictive Behavior: A Study of a HIV-1 Transgenic Rat Model  

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Full Text Available Epidemiological research indicates that drug abuse is prevalent among individuals infected with HIV-1. Evidence from preclinical research also suggests that drugs of abuse exacerbate the progression of neuropathological changes in the HIV-1 infected brain probably through common mechanisms of neuronal injury. The effects of HIV-1 on the efficacy and abuse potential of controlled drugs such as morphine, however, has not been explored. The current study reports that the noninfectious HIV-1 transgenic (HIV-1 Tg rat shows up-regulated expression of the mu opioid receptor (MOR at the transcriptional level and functional supersensitivity to morphine, a MOR agonist. Compared to nontransgenic control rats, the HIV-1 Tg rats also show greater motivation to run in a wheel, a behavior that is known to be associated with increased drug self-administration. These results suggest the potential role of HIV-1 infection in enhancing vulnerability to addiction and this possibility warrants further investigation to better understand the link between HIV-1 infection and the abuse of drugs including opioids.

Sulie L. Chang

2006-01-01

183

Increased neuroinflammatory and arachidonic acid cascade markers, and reduced synaptic proteins, in brain of HIV-1 transgenic rats  

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Full Text Available Abstract Background Cognitive impairment has been reported in human immune deficiency virus-1- (HIV-1- infected patients as well as in HIV-1 transgenic (Tg rats. This impairment has been linked to neuroinflammation, disturbed brain arachidonic acid (AA metabolism, and synapto-dendritic injury. We recently reported upregulated brain AA metabolism in 7- to 9-month-old HIV-1 Tg rats. We hypothesized that these HIV-1 Tg rats also would show upregulated brain inflammatory and AA cascade markers and a deficit of synaptic proteins. Methods We measured protein and mRNA levels of markers of neuroinflammation and the AA cascade, as well as pro-apoptotic factors and synaptic proteins, in brains from 7- to 9-month-old HIV-1 Tg and control rats. Results Compared with control brain, HIV-1 Tg rat brain showed immunoreactivity to glycoprotein 120 and tat HIV-1 viral proteins, and significantly higher protein and mRNA levels of (1 the inflammatory cytokines interleukin-1? and tumor necrosis factor ?, (2 the activated microglial/macrophage marker CD11b, (3 AA cascade enzymes: AA-selective Ca2+-dependent cytosolic phospholipase A2 (cPLA2-IVA, secretory sPLA2-IIA, cyclooxygenase (COX-2, membrane prostaglandin E2 synthase, 5-lipoxygenase (LOX and 15-LOX, cytochrome p450 epoxygenase, and (4 transcription factor NF-?Bp50 DNA binding activity. HIV-1 Tg rat brain also exhibited signs of cell injury, including significantly decreased levels of brain-derived neurotrophic factor (BDNF and drebrin, a marker of post-synaptic excitatory dendritic spines. Expression of Ca2+-independent iPLA2-VIA and COX-1 was unchanged. Conclusions HIV-1 Tg rats show elevated brain markers of neuroinflammation and AA metabolism, with a deficit in several synaptic proteins. These changes are associated with viral proteins and may contribute to cognitive impairment. The HIV-1 Tg rat may be a useful model for understanding progression and treatment of cognitive impairment in HIV-1 patients.

Harry Gaylia

2011-08-01

184

Ankylosing spondylitis in West Africans--evidence for a non-HLA-B27 protective effect.  

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OBJECTIVE: To determine the prevalence of ankylosing spondylitis in the Fula ethnic group in The Gambia, and relate the disease prevalence to the B27 frequency and subtype distribution of that population. METHODS: 215 first degree relatives of 48 B27 positive Fula twin pairs, and 900 adult Fula males were screened for ankylosing spondylitis by clinical and, where appropriate, radiographic means. The B27 prevalence was determined by PCR/sequence specific oligonucleotides on finger prick sample...

Brown, Ma; Jepson, A.; Young, A.; Whittle, Hc; Greenwood, BM; Wordsworth, Bp

1997-01-01

185

Determination of HLA-B27 Subtypes in Iranian Patients with Ankylosing Spondylitis  

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The human leukocyte antigen-B27 is one of the class I molecules of the major histocompatibility complex which is strongly associated with ankylosing spondylitis (AS). The strength of the disease association with B27 varies markedly among racial and ethnic populations. It is an allele family, which constitutes about 31 subtypes, with a considerable geographic and ethnic difference in distribution. It is important to know whether certain subtypes show any preferential association with AS. Becau...

Behrooz Nikbin; Mostafa Moin; Zahra Pourpak; Bita Ansaripour; Aref Amirkhani; Ahmad Reza Jamshidi; Farideh Khosravi; Mazdak Ganjalikhani Hakemi; Ali Akbar Amirzargar; Mahdi Mahmoudi; Mohamad Hossein Nicknam

2008-01-01

186

HIV-1 transgenic rat CD4+ T cells develop decreased CD28 responsiveness and suboptimal Lck tyrosine dephosphorylation following activation  

International Nuclear Information System (INIS)

Impaired CD4+ T cell responses, resulting in dysregulated T-helper 1 (Th1) effector and memory responses, are a common result of HIV-1 infection. These defects are often preceded by decreased expression and function of the ?/? T cell receptor (TCR)-CD3 complex and of co-stimulatory molecules including CD28, resulting in altered T cell proliferation, cytokine secretion and cell survival. We have previously shown that HIV Tg rats have defective development of T cell effector function and generation of specific effector/memory T cell subsets. Here we identify abnormalities in activated HIV-1 Tg rat CD4+ T cells that include decreased pY505 dephosphorylation of Lck (required for Lck activation), decreased CD28 function, reduced expression of the anti-apoptotic molecule Bcl-xL, decreased secretion of the mitogenic lympokine interleukin-2 (IL-2) and increased activation induced apoptosis. These events likely lead to defects in antigen-specific signaling and may help explain the disruption of Th1 responses and the generation of specific effector/memory subsets in transgenic CD4+ T cells

187

Medial prefrontal cortex neuronal activation and synaptic alterations after stress-induced reinstatement of palatable food seeking: a study using c-fos-GFP transgenic female rats.  

Science.gov (United States)

Relapse to maladaptive eating habits during dieting is often provoked by stress and there is evidence for a role of ovarian hormones in stress responses and feeding. We studied the role of these hormones in stress-induced reinstatement of food seeking and medial prefrontal cortex (mPFC) neuronal activation in c-fos-GFP transgenic female rats, which express GFP in strongly activated neurons. Food-restricted ovariectomized or sham-operated c-fos-GFP rats were trained to lever-press for palatable food pellets. Subsequently, lever-pressing was extinguished and reinstatement of food seeking and mPFC neuronal activation was assessed after injections of the pharmacological stressor yohimbine (0.5-2 mg/kg) or pellet priming (1-4 noncontingent pellets). Estrous cycle effects on reinstatement were also assessed in wild-type rats. Yohimbine- and pellet-priming-induced reinstatement was associated with Fos and GFP induction in mPFC; both reinstatement and neuronal activation were minimally affected by ovarian hormones in both c-fos-GFP and wild-type rats. c-fos-GFP transgenic rats were then used to assess glutamatergic synaptic alterations within activated GFP-positive and nonactivated GFP-negative mPFC neurons following yohimbine-induced reinstatement of food seeking. This reinstatement was associated with reduced AMPA receptor/NMDA receptor current ratios and increased paired-pulse facilitation in activated GFP-positive but not GFP-negative neurons. While ovarian hormones do not appear to play a role in stress-induced relapse of food seeking in our rat model, this reinstatement was associated with unique synaptic alterations in strongly activated mPFC neurons. Our paper introduces the c-fos-GFP transgenic rat as a new tool to study unique synaptic changes in activated neurons during behavior. PMID:22723688

Cifani, Carlo; Koya, Eisuke; Navarre, Brittany M; Calu, Donna J; Baumann, Michael H; Marchant, Nathan J; Liu, Qing-Rong; Khuc, Thi; Pickel, James; Lupica, Carl R; Shaham, Yavin; Hope, Bruce T

2012-06-20

188

Bone marrow-derived mesenchymal stem cells expressing the Shh transgene promotes functional recovery after spinal cord injury in rats.  

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Spinal cord injury (SCI) is one of the most disabling diseases. Cell-based gene therapy is becoming a major focus for the treatment of SCI. Bone marrow-derived mesenchymal stem cells (BMSCs) are a promising stem cell type useful for repairing SCI. However, the effects of BMSCs transplants are likely limited because of low transplant survival after SCI. Sonic hedgehog (Shh) is a multifunctional growth factor which can facilitate neuronal and BMSCs survival, promote axonal growth, prevent activation of the astrocyte lineage, and enhance the delivery of neurotrophic factors in BMSCs. However, treatment of SCI with Shh alone also has limited effects on recovery, because the protein is cleared quickly. In this study, we investigated the use of BMSCs overexpressing the Shh transgene (Shh-BMSCs) in the treatment of rats with SCI, which could stably secrete Shh and thereby enhance the effects of BMSCs, in an attempt to combine the advantages of Shh and BMSCs and so to promote functional recovery. After Shh-BMSCs treatment of SCI via the subarachnoid, we detected significantly greater damage recovery compared with that seen in rats treated with phosphate-buffered saline (PBS) and BMSCs. Use of Shh-BMSCs increased the expression and secretion of Shh, basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), improved the behavioral function, enhanced the BMSCs survival, promoted the expression level of neurofilament 200 (NF200), and reduced the expression of glial fibrillary acidic protein (GFAP). Thus, our results indicated that Shh-BMSCs enhanced recovery of neurological function after SCI in rats and could be a potential valuable therapeutic intervention for SCI in humans. PMID:24837681

Jia, Yijia; Wu, Dou; Zhang, Ruiping; Shuang, Weibing; Sun, Jiping; Hao, Haihu; An, Qijun; Liu, Qiang

2014-06-24

189

Circadian dysfunction in P23H rhodopsin transgenic rats: effects of exogenous melatonin.  

Science.gov (United States)

This study focuses on the effects of retinal degeneration on the circadian patterns of P23H rats, as well as on the effect of exogenous melatonin administration. To this end, the body temperature of P23H and Sprague-Dawley rats was continuously monitored and their retinas examined at different stages of degeneration, by means of histological labeling and electroretinogram recordings. Melatonin (2?mg/kg BW/day) was supplied ad libitum throughout the experiment to a subset of animals. The body temperature recordings from wild-type and mutant animals showed no differences in the periodogram and the pattern of the mean waveform. However, a progressive decrease in the relative amplitude of the rhythm (RA), a decline in the coupling strength of the rhythm to environmental zeitgebers (interdaily stability, IS) and increased rhythm fragmentation (intradaily variability, IV) were observed in P23H rats, when compared to wild-type animals. The P23H animals showed a progressive decrease in light-induced retinal responses until reaching 18?months of age. By this age, all photoreceptors had already disappeared, and no responses were found in the EGRs. Exogenous administration of melatonin improved the visual response of P23H rats. In fact, the maximum b-wave recorded at 14?months of age was significantly higher in melatonin-treated P23H rats than in the control animals. Furthermore, the maximum b-wave recorded for P23H rats at the age of 14?months significantly correlated with RA, IS, and IV. This leads us to conclude that vision loss in P23H rats is correlated with a progressive fragmentation of their circadian patterns. Both effects are partially reversed by melatonin administration. PMID:21062354

Lax, Pedro; Otalora, Beatriz Baño; Esquiva, Gema; Rol, María de Los Ángeles; Madrid, Juan Antonio; Cuenca, Nicolás

2011-03-01

190

Combined inhibition of 20-hydroxyeicosatetraenoic acid formation and of epoxyeicosatrienoic acids degradation attenuates hypertension and hypertension-induced end-organ damage in Ren-2 transgenic rats  

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Abstract Recent studies have shown that the renal cytochrome P-450 metabolites of arachidonic acid: the vasoconstrictor 20-hydroxyeicosatetraenoic acid (20-HETE), and the vasodilator epoxyeicosatrienoic acids (EETs) play an important role in the pathophysiology of angiotensin II (ANG II)-dependent forms of hypertension and the associated target organ damage. The present studies were performed in Ren-2 renin transgenic rats (TGR) to evaluate the effects of chronic selective inhibiti...

C?erti?kova? Cha?bova?, Ve?ra; Walkowska, Agnieszka; Kompanowska-jezierska, Elzbieta; Sadowski, Janusz; Kujal, Peter; Vernerova?, Zdena; Van?ourkova?, Zdenka; Kopkan, Libor; Kramer, Herbert J.; Falck, John R.; Imig, John D.; Hammock, Bruce D.; Vane?c?kova?, Ivana; C?ervenka, Lude?k

2010-01-01

191

Imatinib ameliorates renal morphological changes in Cyp1a1-Ren2 transgenic rats with inducible ANG II-dependent malignant hypertension  

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The present study was performed to assess the effects of the platelet-derived growth factor (PDGF) receptor kinase inhibitor imatinib mesylate on the renal morphological changes occurring during the development of malignant hypertension in transgenic rats with inducible expression of the Ren2 gene [TGR(Cyp1a1Ren2)]. Arterial blood pressure was measured by radiotelemetry in male Cyp1a1-Ren2 rats during control conditions and during dietary administration of indole-3-carbinol (I3C; 0.3%) for 14...

Graciano, Miguel L.; Mitchell, Kenneth D.

2011-01-01

192

Cardiac remodeling during and after renin-angiotensin system stimulation in Cyp1a1-Ren2 transgenic rats  

DEFF Research Database (Denmark)

This study investigated renin-angiotensin system (RAS)-induced cardiac remodeling and its reversibility in the presence and absence of high blood pressure (BP) in Cyp1a1-Ren2 transgenic inducible hypertensive rats (IHR). In IHR (pro)renin levels and BP can be dose-dependently titrated by oral administration of indole-3-carbinol (I3C). Young (four-weeks old) and adult (30-weeks old) IHR were fed I3C for four weeks (leading to systolic BP >200 mmHg). RAS-stimulation was stopped and animals were followed-up for a consecutive period. Cardiac function and geometry was determined echocardiographically and the hearts were excised for molecular and immunohistochemical analyses. Echocardiographic studies revealed that four weeks of RAS-stimulation incited a cardiac remodeling process characterized by increased left ventricular (LV) wall thickness, decreased LV volumes, and shortening of the left ventricle. Hypertrophic genes were highly upregulated, whereas in substantial activation a fibrotic response was absent. Four weeks after withdrawal of I3C, (pro)renin levels were normalized in all IHR. While in adult IHR BP returned to normal, hypertension was sustained in young IHR. Despite the latter, myocardial hypertrophy was fully regressed in both young and adult IHR. We conclude that (pro)renin-induced severe hypertension in IHR causes an age-independent fully reversible myocardial concentric hypertrophic remodeling, despite a continued elevated BP in young IHR.

Heijnen, Bart Fj; Pelkmans, Leonie Pj

2013-01-01

193

Establishment of mesenchymal stem cells derived from bone marrow and synovium of transgenic rats expressing dual reporter genes  

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Mesenchymal stem cells (MSCs) are an attractive cell source for regenerative medicine because they can be harvested in a relatively less invasive manner, easily isolated, and expanded with multipotentiality. Bone marrow seems to be the most commonly used tissue as a source for MSCs at present. However, there are emerging reports to describe that MSCs exist in most mesenchymal tissues. We have recently compared in vivo chondrogenic potential in MSCs derived from various mesenchymal tissues and demonstrated that synovium-MSCs and bone marrow-MSCs possessed greater chondrogenic ability than other mesenchymal tissue-derived MSCs. This indicates that those MSCs are promising cellular sources for cartilage regeneration. As the fate of synovium-MSCs is unclear after transplantation, we herein established MSCs using double transgenic rats expressing either Luciferase/GFP or Luciferase/LacZ. The cellular fate of MSCs could be traced by an in vivo luciferase-based luminescent imaging system, and also followed histologically by green fluorescence and by X-gal staining, respectively. Thus, both synovium-MSCs and bone marrow-MSCs expressing Luciferase/GFP or Luciferase/LacZ provide powerful tools not only for cell tracking in vivo but also for histological analysis, leading to a compelling experimental model of cartilage regeneration with cell therapy.

Horie, Masafumi; Sekiya, Ichiro; Muneta, Takeshi; Murakami, Takashi; Kobayashi, Eiji

2008-02-01

194

Transgenic rats reveal functional conservation of regulatory controls between the Fugu isotocin and rat oxytocin?genes  

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We have asked whether comparative genome analysis and rat transgenesis can be used to identify functional regulatory domains in the gene locus encoding the hypothalamic neuropeptides oxytocin (OT) and vasopressin. Isotocin (IT) and vasotocin (VT) are the teleost homologues of these genes. A contiguous stretch of 46 kb spanning the Fugu IT-VT locus has been sequenced, and nine putative genes were found. Unlike the OT and vasopressin genes, which are closely linked in the mammalian genome in a ...

Venkatesh, Byrappa; Si-hoe, San Ling; Murphy, David; Brenner, Sydney

1997-01-01

195

Vitamin A Deficiency and Behavioral and Motor Deficits in the HIV-1 Transgenic Rat  

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The HIV-1 Tg rat model incorporates a non-infectious viral genome that is under similar regulatory control mechanisms in vivo as those that exist with natural infection in humans. Vitamin A (VA) deficiency in humans has been associated with progressive systemic HIV disease and with impaired cognition in rodent models. The effects on of VA deficiency on the development of behavioral abnormalities with HIV infection have not been previously described. In these studies wild type (Wt) and transge...

June, Harry L.; Yang, Andrew Rong Song Tzeng; Bryant, Joseph L.; Jones, Odell; Royal, Walter

2009-01-01

196

Different mechanisms of acute versus long-term antihypertensive effects of soluble epoxide hydrolase inhibition: Studies in Cyp1a1-Ren-2 transgenic rats.  

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Recent studies have shown that the long-term antihypertensive action of soluble epoxide hydrolase inhibition (sEH) in angiotensin-II (AngII)-dependent hypertension might be mediated by the suppression of intrarenal AngII levels. To test this hypothesis, we examined the effects of acute (2 days) and chronic (14 days) sEH inhibition on blood pressure (BP) in transgenic rats with inducible AngII-dependent hypertension. AngII-dependent malignant hypertension was induced by 10 days' dietary administration of indole-3-carbinol (I3C), a natural xenobiotic that activates the mouse renin gene in Cyp1a1-Ren-2 transgenic rats. BP was monitored by radiotelemetry. Acute and chronic sEH inhibition was achieved using cis-4-(4-(3-adamantan-1-yl-ureido)cyclohexyloxy) benzoic acid, given at doses of 0.3, 3, 13, 26, 60 and 130 mg/L in drinking water. At the end of experiments, renal concentrations of epoxyeicosatrienoic acids, their inactive metabolites dihydroxyeicosatrienoic acids and AngII were measured. Acute BP-lowering effects of sEH inhibition in I3C-induced rats was associated with a marked increase in renal epoxyeicosatrienoic acids to dihydroxyeicosatrienoic acids ratio and acute natriuresis. Chronic treatment with cis-4-(4-(3-adamantan-1-yl-ureido)cyclohexyloxy) benzoic acid in I3C-induced rats elicited dose-dependent persistent BP lowering associated with a significant reduction of plasma and kidney AngII levels. Our findings show that the acute BP-lowering effect of sEH inhibition in I3C-induced Cyp1a1-Ren-2 transgenic rats is mediated by a substantial increase in intrarenal epoxyeicosatrienoic acids and their natriuretic action without altering intrarenal renin-angiotensin system activity. Long-term antihypertensive action of cis-4-(4-(3-adamantan-1-yl-ureido)cyclohexyloxy) benzoic acid in I3C-induced Cyp1a1-Ren-2 transgenic rats is mediated mostly by suppression of intrarenal AngII concentration. PMID:25224811

Sporková, Alexandra; Jíchová, Sárka; Husková, Zuzana; Kopkan, Libor; Nishiyama, Akira; Hwang, Sung H; Hammock, Bruce D; Imig, John D; Kompanowska-Jezierska, Elzbieta; Sadowski, Janusz; Kramer, Herbert J; Cervenka, Lud?k

2014-12-01

197

Plasmid-based genetic modification of human bone marrow-derived stromal cells: analysis of cell survival and transgene expression after transplantation in rat spinal cord  

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Full Text Available Abstract Background Bone marrow-derived stromal cells (MSC are attractive targets for ex vivo cell and gene therapy. In this context, we investigated the feasibility of a plasmid-based strategy for genetic modification of human (hMSC with enhanced green fluorescent protein (EGFP and neurotrophin (NT3. Three genetically modified hMSC lines (EGFP, NT3, NT3-EGFP were established and used to study cell survival and transgene expression following transplantation in rat spinal cord. Results First, we demonstrate long-term survival of transplanted hMSC-EGFP cells in rat spinal cord under, but not without, appropriate immune suppression. Next, we examined the stability of EGFP or NT3 transgene expression following transplantation of hMSC-EGFP, hMSC-NT3 and hMSC-NT3-EGFP in rat spinal cord. While in vivo EGFP mRNA and protein expression by transplanted hMSC-EGFP cells was readily detectable at different time points post-transplantation, in vivo NT3 mRNA expression by hMSC-NT3 cells and in vivo EGFP protein expression by hMSC-NT3-EGFP cells was, respectively, undetectable or declined rapidly between day 1 and 7 post-transplantation. Further investigation revealed that the observed in vivo decline of EGFP protein expression by hMSC-NT3-EGFP cells: (i was associated with a decrease in transgenic NT3-EGFP mRNA expression as suggested following laser capture micro-dissection analysis of hMSC-NT3-EGFP cell transplants at day 1 and day 7 post-transplantation, (ii did not occur when hMSC-NT3-EGFP cells were transplanted subcutaneously, and (iii was reversed upon re-establishment of hMSC-NT3-EGFP cell cultures at 2 weeks post-transplantation. Finally, because we observed a slowly progressing tumour growth following transplantation of all our hMSC cell transplants, we here demonstrate that omitting immune suppressive therapy is sufficient to prevent further tumour growth and to eradicate malignant xenogeneic cell transplants. Conclusion In this study, we demonstrate that genetically modified hMSC lines can survive in healthy rat spinal cord over at least 3 weeks by using adequate immune suppression and can serve as vehicles for transgene expression. However, before genetically modified hMSC can potentially be used in a clinical setting to treat spinal cord injuries, more research on standardisation of hMSC culture and genetic modification needs to be done in order to prevent tumour formation and transgene silencing in vivo.

Van Tendeloo Viggo FI

2007-12-01

198

Use of a cryptic splice donor site in the chloramphenicol acetyltransferase (CAT)-SV40 small-t antigen cassette generates alternative transcripts in transgenic rats.  

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The bacterial gene chloramphenicol acetyltransferase (CAT) is a widely used reporter in both in-vitro and in-vivo studies of genetic regulation. We have recently generated novel rat transgenic lines carrying an arylalkylamine N-acetyltransferase (AA-NAT) promoter-reporter construct in which CAT (with associated SV40 small-t antigen sequence) is the reporter. In addition to the predicted transgene transcript (1.9 kb), we identified an abundant 1.5 kb transcript which derives from an alternative splicing event that utilises a cryptic splice donor site located within the CAT gene. The native CAT open reading frame (ORF) is lost in the 1.5 kb transcript, and a western analysis has shown that protein deriving from an aberrant open reading frame is not expressed at detectable levels. PMID:10853270

Burke, Z D; Wells, T; Carter, D A; Baler, R

2000-02-01

199

Sterol regulatory element binding protein 2 overexpression is associated with reduced adipogenesis and ectopic fat accumulation in transgenic spontaneously hypertensive rats.  

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It has been reported that the major function of the sterol regulatory element binding protein 2 (SREBP-2) is to activate preferentially cholesterol biosynthesis in liver and adipose tissue rather than fatty acid synthesis. In the current study, we analyzed the effects of overexpression of human dominant-positive SREBP-2 transgene under control of PEPCK promoter in the spontaneously hypertensive rat (SHR) on lipid and glucose metabolism. Transgenic overexpression of SREBP-2 was associated with significantly higher hepatic triglycerides (20.4+/-0.9 vs. 17.0+/-0.05 micromol/g, Prats when compared to SHR controls. Ectopic fat accumulation was associated with significantly increased serum glucose levels (6.4+/-0.1 vs. 5.9+/-0.1 mmol/l, Prats. These results provide evidence for important role of SREBP-2 in regulation of lipid and glucose metabolism. PMID:24908080

Landa, V; Zídek, V; Mlejnek, P; Simáková, M; Silhavý, J; Trnovská, J; Kazdová, L; Pravenec, M

2014-11-27

200

Dynamics of testicular germ cell apoptosis in normal mice and transgenic mice overexpressing rat androgen-binding protein  

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Full Text Available Abstract The number and type of testicular germ cells undergoing apoptosis in different age groups of mice (from 7 to 360 days of age was determined and compared in age-matched wild type (WT control and in a transgenic (TG mice homozygous to rat androgen binding protein (ABP using flow cytometry. Flow cytometric quantification revealed that the total number of germ cells undergoing apoptosis did not differ significantly in WT and TG mice up to Day 14. From Day 21 to Day 60, the number of germ cells undergoing apoptosis was consistently higher in TG than in WT mice. Starting from Day 90, the number of germ cells undergoing apoptosis in TG mice was lower than controls until Day 360. In 21–60 days old TG mice, spermatogonia, S-Phase cells, and primary spermatocytes are the cell types undergoing apoptosis at significantly greater numbers than those in WT mice. However, starting from day 60, the total number of spermatids undergoing apoptosis was significantly lower in TG mice than in age-matched WT controls. TdT-mediated dUTP-biotin nick end labeling (TUNEL in testicular sections from TG mice of 21 and 30 days of age confirmed the presence of increased numbers of apoptotic germ cells compared to their age matched controls. These data indicate that the continuous presence of greater than physiological concentrations of ABP in the mouse testis has a biphasic effect on the frequency of apoptosis in germ cells. The initial pre-pubertal increase in testicular germ cell apoptosis may result from direct or indirect actions of ABP and is likely to determine the subsequent life-death balance of germ cell populations in TG mice, whereas the subsequent reduction may result from maturation depletion. A wave of apoptosis during the pre-pubertal period is required for normal spermatogenesis to develop, and our data indicate that this apoptotic wave may be regulated by ABP and/or androgens.

Petrusz Peter

2003-06-01

 
 
 
 
201

Transgenic mouse lines expressing rat AH receptor variants - A new animal model for research on AH receptor function and dioxin toxicity mechanisms  

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Han/Wistar (Kuopio; H/W) rats are exceptionally resistant to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) toxicity mainly because of their mutated aryl hydrocarbon receptor (AHR) gene. In H/W rats, altered splicing of the AHR mRNA generates two AHR proteins: deletion (DEL) and insertion (INS) variants, with the INS isoform being predominantly expressed. To gain further insight into their functional properties, cDNAs of these and rat wild-type (rWT) isoform were transferred into C57BL/6J-derived mice by microinjection. The endogenous mouse AHR was eliminated by selective crossing with Ahr-null mice. A single mouse line was obtained for each of the three constructs. The AHR mRNA levels in tissues were generally close to those of C57BL/6 mice in INS and DEL mice and somewhat higher in rWT mice; in testis, however, all 3 constructs exhibited marked overexpression. The transgenic mouse lines were phenotypically normal except for increased testis weight. Induction of drug-metabolizing enzymes by TCDD occurred similarly to that in C57BL/6 mice, but there tended to be a correlation with AHR concentrations, especially in testis. In contrast to C57BL/6 mice, the transgenics did not display any major gender difference in susceptibility to the acute lethality and hepatotoxicity of TCDD; rWT mice were highly sensitive, DEL mice moderately resistant and INS mice highly resistant. Co-expression of mouse AHR and rWT resulted in augmented sensitivity to TCDD and abolished the natural reivity to TCDD and abolished the natural resistance of female C57BL/6 mice, whereas mice co-expressing mouse AHR and INS were resistant. Thus, these transgenic mouse lines provide a novel promising tool for molecular studies on dioxin toxicity and AHR function.

202

A c-fos-monomeric red fluorescent protein 1 fusion transgene is differentially expressed in rat forebrain and brainstem after chronic dehydration and rehydration.  

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We have previously shown that an acute osmotic stimulation induces the expression of a c-fos and monomeric red fluorescent protein 1 (mRFP1) fusion transgene in osmosensitive rat brain areas, including the supraoptic (SON) and paraventricular nuclei (PVN). However, the effects of chronic stimuli, such as dehydration, have not been investigated. In the present study, the expression patterns of the c-fos-mRFP1 fusion gene in the forebrain and the brainstem of male and female transgenic rats were studied in seven experimental groups: ad lib. water (euhydration), water deprivation for 12, 24 or 48 h (dehydration) and water deprivation for 46 h + ad lib. water for 2, 6 or 12 h (rehydration). The number of cells that express nuclear mRFP1 fluorescence was quantified in the hypothalamus, the circumventricular organs and the brainstem. Compared to the euhydrated state, the number of transgene expressing cells significantly increased in all forebrain areas and in the rostral ventrolateral medulla after dehydration and 2 h of rehydration. In the nucleus of the solitary tract and area postrema, the number of mRFP1 fluorescent cells was markedly increased after 2 h of rehydration. Although the number of mRFP1 fluorescent cells in the organum vasculosum laminae terminalis, median preoptic nucleus and subfornical organ remained significantly increased after 6 h of rehydration, reaching control levels after 12 h of rehydration, the number of mRFP1 fluorescent cells in the SON and the PVN reached control levels after 6 h of rehydration. There were no significant differences between male and female rats. These results show that the expression of the c-fos-mRFP1 fusion gene changes in the forebrain and the brainstem not only after acute osmotic stimulation, but also after chronic osmotic stimulation. Interestingly, these studies reveal the differential activation of different neuronal groups over the time course of dehydration and rehydration. PMID:23350545

Yoshimura, M; Ohkubo, J; Katoh, A; Ohno, M; Ishikura, T; Kakuma, T; Yoshimatsu, H; Murphy, D; Ueta, Y

2013-05-01

203

Novel transgenic mouse models develop retinal changes associated with early diabetic retinopathy similar to those observed in rats with diabetes mellitus.  

Science.gov (United States)

Retinal capillary pericyte degeneration has been linked to aldose reductase (AR) activity in diabetic retinopathy (DR). Since the development of DR in mice and rats has been reported to differ and that this may be linked to differences in retinal sorbitol levels, we have established new murine models of early onset diabetes mellitus as tools for investigating the role of AR in DR. Transgenic diabetic mouse models were developed by crossbreeding diabetic C57BL/6-Ins2(Akita)/J (AK) with transgenic C57BL mice expressing green fluorescent protein (GFP), human aldose reductase (hAR) or both in vascular tissues containing smooth muscle actin-? (SMAA). Changes in retinal sorbitol levels were determined by HPLC while changes of growth factors and signaling were investigated by Western Blots. Retinal vascular changes were quantitatively analyzed on elastase-digestion flat mounts. Results show that sorbitol levels were higher in neural retinas of diabetic AK-SMAA-GFP-hAR compared to AK-SMAA-GFP mice. AK-SMAA-GFP-hAR mice showed induction of the retinal growth factors VEGF, IGF-1, bFGF and TGF?, as well as signaling changes in P-Akt, P-SAPK/JNK, and P-44/42 MAPK. Increased loss of nuclei per capillary length and a significant increase in the percentage of acellular capillaries presented in 18 week old AK-SMAA-GFP-hAR mice. These changes are similar to those observed in streptozotocin-induced diabetic rats. Retinal changes in both mice and rats were prevented by inhibition of AR. These studies confirm that the increased expression of AR in mice results in the development of retinal changes associated with the early stages of DR that are similar to those observed in rats. PMID:24370601

Guo, Changmei; Zhang, Zifeng; Zhang, Peng; Makita, Jun; Kawada, Hiroyoshi; Blessing, Karen; Kador, Peter F

2014-02-01

204

Adeno-associated viral vector serotypes 1 and 5 targeted to the neonatal rat and pig striatum induce widespread transgene expression in the forebrain  

DEFF Research Database (Denmark)

Viral vector-mediated gene transfer has emerged as a powerful means to target transgene expression in the central nervous system. Here we characterized the efficacy of serotypes 1 and 5 recombinant adeno-associated virus (rAAV) vectors encoding green fluorescent protein (GFP) after stereotaxic delivery to the neonatal rat and minipig striatum. The efficiency of GFP expression and the phenotype of GFP-positive cells were assessed within the forebrain at different time points up to 12 months after surgery. Both rAAV1-GFP and rAAV5-GFP delivery resulted in transduction of the striatum as well as striatal input and output areas, including large parts of the cortex. In both species, rAAV5 resulted in a more widespread transgene expression compared to rAAV1. In neonatal rats, rAAV5 also transduced several other areas such as the olfactory bulbs, hippocampus, and septum. Phenotypic analysis of the GFP-positive cells, performed using immunohistochemistry and confocal microscopy, showed that most of the GFP-positive cells by either serotype were NeuN-positive neuronal profiles. The rAAV5 vector further displayed the ability to transduce non-neuronal cell types in both rats and pigs, albeit at a low frequency. Our results show that striatal delivery of rAAV5 vectors in the neonatal brain represents a useful tool to express genes of interest both in the basal ganglia and the neocortex. Furthermore, we apply, for the first time, viral vector-mediated gene transfer to the pig brain providing the opportunity to study effects of genetic manipulation in this non-primate large animal species. Finally, we generated an atlas of the Göttingen minipig brain for guiding future studies in this large animal species.

Kornum, Birgitte R; Stott, Simon R W

2010-01-01

205

Combined renin inhibition/(pro)renin receptor blockade in diabetic retinopathy--a study in transgenic (mREN2)27 rats.  

Science.gov (United States)

Dysfunction of renin-angiotensin system (RAS) contributes to the pathogenesis of diabetic retinopathy (DR). Prorenin, the precursor of renin is highly elevated in ocular fluid of diabetic patients with proliferative retinopathy. Prorenin may exert local effects in the eye by binding to the so-called (pro)renin receptor ((P)RR). Here we investigated the combined effects of the renin inhibitor aliskiren and the putative (P)RR blocker handle-region peptide (HRP) on diabetic retinopathy in streptozotocin (STZ)-induced diabetic transgenic (mRen2)27 rats (a model with high plasma prorenin levels) as well as prorenin stimulated cytokine expression in cultured Müller cells. Adult (mRen2)27 rats were randomly divided into the following groups: (1) non-diabetic; (2) diabetic treated with vehicle; (3) diabetic treated with aliskiren (10 mg/kg per day); and (4) diabetic treated with aliskiren+HRP (1 mg/kg per day). Age-matched non-diabetic wildtype Sprague-Dawley rats were used as control. Drugs were administered by osmotic minipumps for three weeks. Transgenic (mRen2)27 rat retinas showed increased apoptotic cell death of both inner retinal neurons and photoreceptors, increased loss of capillaries, as well as increased expression of inflammatory cytokines. These pathological changes were further exacerbated by diabetes. Aliskiren treatment of diabetic (mRen2)27 rats prevented retinal gliosis, and reduced retinal apoptotic cell death, acellular capillaries and the expression of inflammatory cytokines. HRP on top of aliskiren did not provide additional protection. In cultured Müller cells, prorenin significantly increased the expression levels of IL-1? and TNF-?, and this was completely blocked by aliskiren or HRP, their combination, (P)RR siRNA and the AT1R blocker losartan, suggesting that these effects entirely depended on Ang II generation by (P)RR-bound prorenin. In conclusion, the lack of effect of HRP on top of aliskiren, and the Ang II-dependency of the ocular effects of prorenin in vitro, argue against the combined application of (P)RR blockade and renin inhibition in diabetic retinopathy. PMID:24968134

Batenburg, Wendy W; Verma, Amrisha; Wang, Yunyang; Zhu, Ping; van den Heuvel, Mieke; van Veghel, Richard; Danser, A H Jan; Li, Qiuhong

2014-01-01

206

Transgene therapy for rat anti-Thy1.1 glomerulonephritis via mesangial cell vector with a polyethylenimine/decorin nanocomplex  

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Polyethylenimine (PEI), a cationic polymer, is one of the most efficient non-viral vectors for transgene therapy. Decorin (DCN), a leucine-rich proteoglycan secreted by glomerular mesangial cells (MC), is a promising anti-fibrotic agent for the treatment of glomerulonephritis. In this study, we used PEI-DCN nanocomplexes with different N/P ratios to transfect MC in vitro and deliver the MC vector with PEI-DCN expressing into rat anti-Thy1.1 nephritis kidney tissue via injection into the left renal artery in vivo. The PEI-plasmid DNA complex at N/P 20 had the highest level of transfection efficiency and the lowest level of cytotoxicity in cultured MC. Following injection, the ex vivo gene was transferred successfully into the glomeruli of the rat anti-Thy1.1 nephritis model by the MC vector with the PEI-DCN complex. The exogenous MC with DCN expression was located mainly in the mesangium and the glomerular capillary. Over-expression of DCN in diseased glomeruli could result in the inhibition of collagen IV deposition and MC proliferation. The pathological changes of rat nephritis were alleviated following injection of the vector. These findings demonstrate that the DCN gene delivered by the PEI-DNA nanocomplex with the MC vector is a promising therapeutic method for the treatment of glomerulonephritis.

Sun, Jian-Yong; Sun, Yu; Wu, Hui-Juan; Zhang, Hong-Xia; Zhao, Zhong-Hua; Chen, Qi; Zhang, Zhi-Gang

2012-08-01

207

Genetic targeting: the serotonin N-acetyltransferase promoter imparts circadian expression selectively in the pineal gland and retina of transgenic rats.  

Science.gov (United States)

The arylalkylamine N-acetyltransferase (AA-NAT) gene is strongly expressed in the rat primarily in the pineal gland; low levels of expression are also found in the retina. AA-NAT catalyzes the key regulatory step controlling rhythmic melatonin output: the acetylation of serotonin. In the rat, the AA-NAT gene is expressed at night. This is controlled partly by cyclic AMP (cAMP) acting through a composite cAMP-responsive element-CCAAT site located upstream of the transcription start point. In the present study, we have extended our previous in vitro findings and found that additional elements in the 5' flanking region and first intron play an important role in the regulation of the AA-NAT gene. This led us to test the influence of an AA-NAT 5' flanking segment on the expression of the bacterial chloramphenicol acetyltransferase gene in a rat transgenic model. The results of this study clearly demonstrate that the segment of the AA-NAT gene that encompasses the minimal promoter and the first intron is able to confer the highly specific pineal/retinal and time-of-day patterns of AA-NAT gene expression. This advance also provides a tool that selectively targets genetic expression to pinealocytes and retinal photoreceptors, providing new experimental opportunities to probe gene expression in these tissues. PMID:10501177

Burke, Z; Wells, T; Carter, D; Klein, D; Baler, R

1999-10-01

208

Expression of the c-fos-monomeric red fluorescent protein 1 fusion gene in the spinal cord and the hypothalamic paraventricular nucleus in transgenic rats after nociceptive stimulation.  

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We generated transgenic rats expressing the c-fos and monomeric red fluorescent protein 1 (mRFP1) fusion gene in the central nervous system after adequate stimulation. In the present study, the time-course of the induction patterns of mRFP1 fluorescence in the spinal cord and the paraventricular nucleus (PVN) was compared with that of Fos-like immunoreactivity (LI) within 24 h after subcutaneous (s.c.) injection of 0.9% saline and 5% formalin in both hind paws. Control rats were not treated. In the control and saline/formalin injected rats, scattered mRFP1 fluorescence in the spinal cord and the PVN was observed at 0 min, though there was little Fos-LI in the same region. The mRFP1 fluorescence in the spinal cord and the PVN was increased at 3h after formalin. On the other hand, the changes of Fos-LI in the spinal cord and the PVN were relatively shorter than those of the mRFP1 fluorescence after formalin. These results suggest that the c-fos-mRFP1 fusion gene expression is slightly upregulated in normal conditions and nociceptive stimulation-induced induction of the fusion gene may be maintained longer than the endogenous c-fos gene expression in the spinal cord and the PVN. Next, nocifensive behavior and mRFP1 fluorescence and Fos-LI in the spinal cord and the PVN after s.c. injection of formalin, 4?-phorbol 12,13-didecanoate (4?-PDD) and saline were compared. Although the 4?-PDD injected rats seldom displayed nocifensive behaviors like s.c. saline injection, 4?-PDD injection caused mRFP1 fluorescence and Fos-LI significantly in the spinal cord and the PVN unlike s.c. saline injection. PMID:22960202

Ishikura, Toru; Suzuki, Hitoshi; Yoshimura, Mitsuhiro; Ohkubo, Jun-ichi; Katoh, Akiko; Ohbuchi, Toyoaki; Ohno, Motoko; Fujihara, Hiroaki; Kawasaki, Makoto; Ohnishi, Hideo; Nakamura, Toshitaka; Ueta, Yoichi

2012-10-15

209

Occurrence of ankylosing spondylitis and multiple sclerosis-like syndrome in a HLA-B27 positive patient.  

Science.gov (United States)

Occurrence of multiple sclerosis (MS) in patients with ankylosing spondylitis (AS) has been reported in isolated cases. We describe a white 33-year-old male with a definite familial HLAB27 positive AS and MS-like syndrome. The patient developed acute onset of gait difficulty, postural unsteadiness, dysarthria and right side weakness that resolved within 1 month; after 6 months he presented right-sided face sensory loss, disappeared after 2 weeks. Brain and cervical MRI was performed twice and showed disseminated lesions in space (multiple foci of increased signal intensity in the periventricular white matter, in the corpus callosum, in the hypothalamus, in the brainstem and in the cervical spinal cord) and in time (a new enhancing lesion >3 months after the onset of the clinical event). Visual evoked potentials were markedly altered. Cerebrospinal fluid examination was negative for intrathecal production of oligoclonal bands. Differential diagnosis was considered and other pathologies were excluded. PMID:19444380

Mignarri, Andrea; Dotti, Maria Teresa; Battisti, Carla; Vallone, Ignazio; Federico, Antonio

2009-08-01

210

Transgenic tea  

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Like most of the important crop plants of the world, transgenic technology has also been extended to tea. Both biolistic and Agrobacterium mediated transformation methods have been employed to transform explants like leaves and somatic embryos. While gusand nptil genes were used to optimize parameters and develop protocols for transgenic production, plants expressing stress tolerance genes (osmotin) have also been produced. These methods have opened a whole new era for developing tea plants a...

Bhattacharya, Amita; Saini, U.; Ahuja, P. S.

2006-01-01

211

Resveratrol induces mitochondrial biogenesis and ameliorates Ang II-induced cardiac remodeling in transgenic rats harboring human renin and angiotensinogen genes.  

Science.gov (United States)

There is compelling evidence to indicate an important role for increased local renin-angiotensin system activity in the pathogenesis of cardiac hypertrophy and heart failure. Resveratrol is a natural polyphenol that activates SIRT1, a novel cardioprotective and longevity factor having NAD(+)-dependent histone deacetylase activity. We tested the hypothesis whether resveratrol could prevent from angiotensin II (Ang II)-induced cardiovascular damage. Four-week-old double transgenic rats harboring human renin and human angiotensinogen genes (dTGR) were treated for 4 weeks either with SIRT1 activator resveratrol or SIRT1 inhibitor nicotinamide. Untreated dTGR and their normotensive Sprague-Dawley control rats (SD) received vehicle. Untreated dTGR developed severe hypertension as well as cardiac hypertrophy, and showed pronounced cardiovascular mortality compared with normotensive SD rats. Resveratrol slightly but significantly decreased blood pressure, ameliorated cardiac hypertrophy and prevented completely Ang II-induced mortality, whereas nicotinamide increased blood pressure without significantly influencing cardiac hypertrophy or survival. Resveratrol decreased cardiac ANP mRNA expression and induced cardiac mRNA expressions of mitochondrial biogenesis markers peroxisome proliferator-activated receptor-gamma coactivator (PGC-1alpha), mitochondrial transcription factor (Tfam), nuclear respiratory factor 1 (NRF-1) and cytochrome c oxidase subunit 4 (cox4). Resveratrol dose-dependently increased SIRT1 activity in vitro. Our findings suggest that the beneficial effects of SIRT1 activator resveratrol on Ang II-induced cardiac remodeling are mediated by blood pressure-dependent pathways and are linked to increased mitochondrial biogenesis. PMID:20429690

Biala, Agnieszka; Tauriainen, Eveliina; Siltanen, Antti; Shi, Jin; Merasto, Saara; Louhelainen, Marjut; Martonen, Essi; Finckenberg, Piet; Muller, Dominik N; Mervaala, Eero

2010-06-01

212

Fluorescent visualisation of the hypothalamic oxytocin neurones activated by cholecystokinin-8 in rats expressing c-fos-enhanced green fluorescent protein and oxytocin-monomeric red fluorescent protein 1 fusion transgenes.  

Science.gov (United States)

The up-regulation of c-fos gene expression is widely used as a marker of neuronal activation elicited by various stimuli. Anatomically precise observation of c-fos gene products can be achieved at the RNA level by in situ hybridisation or at the protein level by immunocytochemistry. Both of these methods are time and labour intensive. We have developed a novel transgenic rat system that enables the trivial visualisation of c-fos expression using an enhanced green fluorescent protein (eGFP) tag. These rats express a transgene consisting of c-fos gene regulatory sequences that drive the expression of a c-fos-eGFP fusion protein. In c-fos-eGFP transgenic rats, robust nuclear eGFP fluorescence was observed in osmosensitive brain regions 90 min after i.p. administration of hypertonic saline. Nuclear eGFP fluorescence was also observed in the supraoptic nucleus (SON) and paraventricular nucleus (PVN) 90 min after i.p. administration of cholecystokinin (CCK)-8, which selectively activates oxytocin (OXT)-secreting neurones in the hypothalamus. In double transgenic rats that express c-fos-eGFP and an OXT-monomeric red fluorescent protein 1 (mRFP1) fusion gene, almost all mRFP1-positive neurones in the SON and PVN expressed nuclear eGFP fluorescence 90 min after i.p. administration of CCK-8. It is possible that not only a plane image, but also three-dimensional reconstruction image may identify cytoplasmic vesicles in an activated neurone at the same time. PMID:24730419

Katoh, A; Shoguchi, K; Matsuoka, H; Yoshimura, M; Ohkubo, J-I; Matsuura, T; Maruyama, T; Ishikura, T; Aritomi, T; Fujihara, H; Hashimoto, H; Suzuki, H; Murphy, D; Ueta, Y

2014-05-01

213

Mutation spectrum of 1,2-dibromo-3-chloropropane, an endocrine disruptor, in the lacI transgenic Big Blue Rat2 fibroblast cell line.  

Science.gov (United States)

1,2-Dibromo-3-chloropropane (DBCP), a soil fumigant against nematodes, has been extensively studied for genotoxicity, carcinogenicity and damage to male reproduction-related organs, as a possible endocrine disruptor. However, the precise mechanisms involved in DBCP-induced mutagenesis and carcinogenesis are as yet unknown. Thus, in this study the mutagenicity and mutation spectrum of DBCP was determined using the lacI transgenic Big Blue Rat2 fibroblast cell line. In determining the optimal concentration of DBCP in Big Blue Rat2 fibroblast cells, the 50% inhibition concentration was calculated to be 0.75 mM. When cells were exposed to DBCP concentrations of 0.21, 0.39 and 0.75 mM, the respective relative survival rates were approximately 80, 70 and 50%. The mean mutant frequencies (MFs) (x 10(-5), +/- SEM) of the medium and 1% DMSO solvent controls were determined as 6.43 +/- 0.616 and 5.28 +/- 1.086, respectively. The MFs (x 10(-5), +/- SEM) of cells exposed to 0.21, 0.39 and 0.75 mM DBCP were 8.09 +/- 1.02, 10.86 +/- 2.17 and 12.26 +/- 0.79, respectively, with a dose-dependent effect (ANOVA, P = 0.007). Moreover, MF values for the 0.75 and 0.39 mM DBCP-treated groups were statistically significant (ANOVA, P DBCP-induced groups. Among 31 single base pair substitutions, 25 (62.5%) occurred at G:C base pairs, while six (15%) were at A:T base pairs. The predominant mutation was G:C-->A:T transitions (16/40, 40%), followed by G:C-->T:A transversions (9/40, 22.5%). We conclude that DBCP is a possible base substitution mutagen, especially at guanine bases. PMID:12110625

Ryu, Jae-Chun; Kim, Youn-Jung; Chai, Young-Gyu

2002-07-01

214

Effects of irbesartan and bosentan on the blood pressure and adrenal zona glomerulosa function in heterozygous transgenic TGR[mREN2]27 rats.  

Science.gov (United States)

The role of angiotensin-II (Ang-II) and endothelin-1 (ET-1) in the development of hypertension and zona glomerulosa (ZG) hyperfunction in the transgenic rat strain TGR[mREN2]27 (TGR) has been investigated. Male heterozygous TGR were given per os for 4 weeks the Ang-II ATI receptor antagonist irbesartan (50 mg/kg x day) or the mixed ETA/ETB receptor antagonist bosentan (100 mg/kg x day). A group of TGR received a placebo gavage. Irbesartan lowered blood pressure (BP), while bosentan was ineffective. Conversely, both antagonists decreased plasma aldosterone concentration, the volume of ZG and its parenchymal cells, and in vitro aldosterone secretion by capsule-ZG preparations. Collectively, our results allow us to conclude that (i) only Ang-II is involved in the genesis of hypertension in TGR, while both endogenous Ang-II and ET-1 play a role in the genesis of ZG hyperfunction; and (ii) hyperaldosteronism does not contribute significantly to the development of hypertension in TGR. PMID:10993119

Andreis, P G; Rebuffat, P; Neri, G; Rossi, G P; Nussdorfer, G G

2000-06-23

215

Targeted neuronal nitric oxide synthase transgene delivery into stellate neurons reverses impaired intracellular calcium transients in prehypertensive rats.  

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Hypertension is associated with the early onset of cardiac sympathetic hyperresponsiveness and enhanced intracellular Ca2+concentration [Ca2+]i in sympathetic neurons from both prehypertensive and hypertensive, spontaneously hypertensive rats (SHRs). Oxidative stress is a hallmark of hypertension, therefore, we tested the hypothesis that the inhibitory action of the nitric oxide-cGMP pathway on [Ca2+]i transients is impaired in cardiac sympathetic neurons from the SHR. Stellate ganglia were i...

Li, D.; Nikiforova, N.; Lu, C-j; Wannop, K.; Mcmenamin, M.; Lee, C-w; Buckler, Kj; Paterson, Dj

2013-01-01

216

Transgenic Animals.  

Science.gov (United States)

Describes three methods and their advantages and disadvantages for introducing genes into animals. Discusses the predictability and tissue-specificity of the injected genes. Outlines the applications of transgenic technology for studying gene expression, the early stages of mammalian development, mutations, and the molecular nature of chromosomes.…

Jaenisch, Rudolf

1988-01-01

217

Nogo-A-deficient transgenic rats show deficits in higher cognitive functions, decreased anxiety and altered circadian activity patterns  

Directory of Open Access Journals (Sweden)

Full Text Available Decreased levels of Nogo-A dependent signaling have been shown to affect behavior and cognitive functions. In Nogo-A knockout and knock-down laboratory rodents, behavioral alterations were observed, possibly corresponding with human neuropsychiatric diseases of neurodevelopmental origin, particularly schizophrenia. This study offers further insight into behavioral manifestations of Nogo-A knockdown in laboratory rats, focusing on spatial and non-spatial cognition, anxiety levels, circadian rhythmicity and activity patterns. Demonstrated is an impairment of cognitive functions and behavioral flexibility in a spatial active avoidance task, while non-spatial memory in a step-through avoidance task was spared. No signs of anhedonia, typical for schizophrenic patients, were observed in the animals. Some measures indicated lower anxiety levels in the Nogo-A deficient group. Circadian rhythmicity in locomotor activity was preserved in the Nogo-A-knockout rats and their circadian period (tau did not differ from controls. However, daily activity patterns were slightly altered in the knockdown animals. We conclude that a reduction of Nogo-A levels induces changes in CNS development, manifested as subtle alterations in cognitive functions, emotionality and activity patterns.

Tomas Petrasek

2014-03-01

218

The vestibulo- and preposito-cerebellar cholinergic neurons of a ChAT-tdTomato transgenic rat exhibit heterogeneous firing properties and the expression of various neurotransmitter receptors.  

Science.gov (United States)

Cerebellar function is regulated by cholinergic mossy fiber inputs that are primarily derived from the medial vestibular nucleus (MVN) and prepositus hypoglossi nucleus (PHN). In contrast to the growing evidence surrounding cholinergic transmission and its functional significance in the cerebellum, the intrinsic and synaptic properties of cholinergic projection neurons (ChPNs) have not been clarified. In this study, we generated choline acetyltransferase (ChAT)-tdTomato transgenic rats, which specifically express the fluorescent protein tdTomato in cholinergic neurons, and used them to investigate the response properties of ChPNs identified via retrograde labeling using whole-cell recordings in brainstem slices. In response to current pulses, ChPNs exhibited two afterhyperpolarisation (AHP) profiles and three firing patterns; the predominant AHP and firing properties differed between the MVN and PHN. Morphologically, the ChPNs were separated into two types based on their soma size and dendritic extensions. Analyses of the firing responses to time-varying sinusoidal current stimuli revealed that ChPNs exhibited different firing modes depending on the input frequencies. The maximum frequencies in which each firing mode was observed were different between the neurons that exhibited distinct firing patterns. Analyses of the current responses to the application of neurotransmitter receptor agonists revealed that the ChPNs expressed (i) AMPA- and NMDA-type glutamate receptors, (ii) GABAA and glycine receptors, and (iii) muscarinic and nicotinic acetylcholine receptors. The current responses mediated by these receptors of MVN ChPNs were not different from those of PHN ChPNs. These findings suggest that ChPNs receive various synaptic inputs and encode those inputs appropriately across different frequencies. PMID:24593297

Zhang, Yue; Kaneko, Ryosuke; Yanagawa, Yuchio; Saito, Yasuhiko

2014-04-01

219

Prostate cancer in a transgenic mouse.  

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Progress toward understanding the biology of prostate cancer has been slow due to the few animal research models available to study the spectrum of this uniquely human disease. To develop an animal model for prostate cancer, several lines of transgenic mice were generated by using the prostate-specific rat probasin promoter to derive expression of the simian virus 40 large tumor antigen-coding region. Mice expressing high levels of the transgene display progressive forms of prostatic disease ...

Greenberg, N. M.; Demayo, F.; Finegold, M. J.; Medina, D.; Tilley, W. D.; Aspinall, J. O.; Cunha, G. R.; Donjacour, A. A.; Matusik, R. J.; Rosen, J. M.

1995-01-01

220

Imaging Transgene Activity  

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The successful translation of gene therapy for clinical application will require the assessment of transgene activity as a measure of the biological function of a therapeutic transgene. While current imaging permits the non-invasive detection of transgene expression, the critical need for quantitative imaging of the action of the expressed transgene has not been met. Magnetic resonance spectroscopic imaging (MRSI) was applied to quantitatively delineate both the concentration and activity of ...

Gade, Terence P. F.; Koutcher, Jason A.; Spees, William M.; Beattie, Bradley J.; Ponomarev, Vladimir; Doubrovin, Michael; Buchanan, Ian M.; Beresten, Tatiana; Zakian, Kristen L.; Le, H. Carl; Tong, William P.; Mayer-kuckuk, Philipp; Blasberg, Ronald G.; Gelovani, Juri G.

2008-01-01

 
 
 
 
221

PRODUCTION OF TRANSGENIC INSECTS  

Science.gov (United States)

Transformation may be defined as insertion of exogenous genes into the germ-line of a novel species, so that the transgene is inherited as part of the transgenic organism’s genome. Examples of transgenic organisms are: Example 1: “Roundup Ready” Soybean (or other plant) Bacterial enzyme gene i...

222

Plasmid-based genetic modification of human bone marrow-derived stromal cells: analysis of cell survival and transgene expression after transplantation in rat spinal cord  

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Abstract Background Bone marrow-derived stromal cells (MSC) are attractive targets for ex vivo cell and gene therapy. In this context, we investigated the feasibility of a plasmid-based strategy for genetic modification of human (h)MSC with enhanced green fluorescent protein (EGFP) and neurotrophin (NT)3. Three genetically modified hMSC lines (EGFP, NT3, NT3-EGFP) were established and used to study cell survival and transgene expression following transplantation in r...

Fi, Tendeloo Viggo; Haese Patrick, D.; Vermeulen Katrien; Chatterjee Shyama; Spaepen Gie; Daans Jasmijn; Ronsyn Mark W; Marck Eric Van; Ysebaert Dirk; Berneman Zwi N; Jorens Philippe G; Ponsaerts Peter

2007-01-01

223

Expression and function of HLA-B27 in lipid-linked form: implications for cytotoxic T lymphocyte-induced apoptosis signal transduction.  

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Major histocompatibility complex (MHC) class I antigen-restricted cytotoxic T lymphocytes (CTL) kill their target cells not only by inducing irreversible membrane damage but also by triggering a programmed suicide cascade (apoptosis) in target cells. Recent evidence suggests that MHC class I antigens are involved in apoptosis signal transduction in T cells. Therefore, it is possible that MHC class I antigens are also responsible for CTL-induced signal transduction in target cells leading to a...

Gao, Xm; Quinn, Cl; Bell, JI; Mcmichael, Aj

1993-01-01

224

HLA-B27 i HLA-DR w prognozowaniu przebiegu m?odzie?czego idiopatycznego zapalenia stawów o pocz?tku uogólnionym  

Directory of Open Access Journals (Sweden)

Full Text Available Przebieg m?odzie?czego idiopatycznego zapalenia stawówo pocz?tku uogólnionym (UMIZS jest zró?nicowany. W artykuleoceniono wp?yw antygenu B27 i serii HLA klasy II – DR na post?pchoroby, ci??ko?? objawów pozastawowych oraz rozwój amyloidozyu 47 chorych z UMIZS z wieloletnim czasem trwania choroby(?rednio 18 ±7,4 roku. G?ównymi parametrami klinicznymi istotniewp?ywaj?cymi na losy chorych, które poddano analizie, by?yzmiany radiologiczne, wydolno?? czynno?ciowa, ci??ko?? zmianstawowych oraz rozwój amyloidozy. U ka?dego pacjenta okre?lonorównie? ci??ko?? objawów uogólnienia, które obserwowanow czasie d?ugoletniej choroby.Wykazano istotn? zale?no?? pomi?dzy obecno?ci? HLA-DR4 a rozwojemzmian radiologicznych w uk?adzie ruchu. HLA-DR4 stwierdzonoznamiennie cz??ciej u chorych ze znacznymi zmianamiradiologicznymi w porównaniu z grup? kontroln? (73,7 vs 23,6%,p < 0,0001, a tak?e w stosunku do chorych bez tych zmian (73,7vs 25%, p < 0,05 (ryc. 4. Antygen DR4 wykrywano równie? znamienniecz??ciej w grupie chorych z najbardziej zaawansowanymizmianami stawowymi w porównaniu z grup? kontroln? (63 vs24%, p < 0,001 (ryc. 2. Istotne powi?zanie z wyst?powaniemobjawów uogólnienia dotyczy?o natomiast HLA-DR3. HLA-DR3istotnie cz??ciej w porównaniu z grup? kontroln? stwierdzono u chorych z objawami uogólnienia wyst?puj?cymi nie tylko napocz?tku choroby, ale równie? w jej dalszym przebiegu (76,2 vs23,6%, p < 0,001, jak i pomi?dzy grup? pacjentów z objawamiuogólnienia ograniczonymi do 2 lat choroby a podgrup? chorychz objawami nawracaj?cymi w dalszym przebiegu choroby (22,7vs 76,2%, p < 0,001 (ryc. 1.Cz?sto?? typowanych antygenów w wyodr?bnionych podgrupach,zarówno w zale?no?ci od wydolno?ci uk?adu ruchu, jak i rozwojuamyloidozy nie ró?ni?a si? istotnie statystycznie.Typowanie HLA mo?e by? pomocne w prognozowaniu dalszegoprzebiegu UMIZS, szczególnie pomaga w identyfikacji chorych,u których dochodzi do zmian destrukcyjnych oraz nawracania objawówuogólnienia w dalszym przebiegu choroby.

Agnieszka Gazda

2011-02-01

225

A hypothesis for the HLA-B27 immune dysregulation in spondyloarthropathy: contributions from enteric organisms, B27 structure, peptides bound by B27, and convergent evolution.  

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Several human rheumatic diseases occur predominantly in persons who carry the histocompatibility (HLA) class I allele B27. They have also been related to Gram-negative enteric microorganisms. In addition, the recent recovery of peptides bound to B27 has allowed an understanding of the structural requirements for their binding. Using the accumulated data base of protein sequences, we have tested a series of hypotheses. First, we have asked whether the primary amino acid sequence of the hyperva...

Scofield, R. H.; Warren, W. L.; Koelsch, G.; Harley, J. B.

1993-01-01

226

Electrophysiological effects of kainic acid on vasopressin-enhanced green fluorescent protein and oxytocin-monomeric red fluorescent protein 1 neurones isolated from the supraoptic nucleus in transgenic rats.  

Science.gov (United States)

The supraoptic nucleus (SON) contains two types of magnocellular neurosecretory cells: arginine vasopressin (AVP)-producing and oxytocin (OXT)-producing cells. We recently generated and characterised two transgenic rat lines: one expressing an AVP-enhanced green fluorescent protein (eGFP) and the other expressing an OXT-monomeric red fluorescent protein 1 (mRFP1). These transgenic rats enable the visualisation of AVP or OXT neurones in the SON. In the present study, we compared the electrophysiological responses of AVP-eGFP and OXT-mRFP1 neurones to glutamic acid in SON primary cultures. Glutamate mediates fast synaptic transmission through three classes of ionotrophic receptors: the NMDA, AMPA and kainate receptors. We investigated the contributions of the three classes of ionotrophic receptors in glutamate-induced currents. Three different antagonists were used, each predominantly selective for one of the classes of ionotrophic receptor. Next, we focused on the kainate receptors (KARs). We examined the electrophysiological effects of kainic acid (KA) on AVP-eGFP and OXT-mRFP1 neurones. In current clamp mode, KA induced depolarisation and increased firing rates. These KA-induced responses were inhibited by the non-NMDA ionotrophic receptor antagonist 6-cyano-7-nitroquinoxaline-2,3(1H4H)-dione in both AVP-eGFP and OXT-mRFP1 neurones. In voltage clamp mode, the application of KA evoked inward currents in a dose-dependent manner. The KA-induced currents were significantly larger in OXT-mRFP1 neurones than in AVP-eGFP neurones. This significant difference in KA-induced currents was abolished by the GluK1-containing KAR antagonist UBP302. At high concentrations (250-500 ?m), the specific GluK1-containing KAR agonist (RS)-2-amino-3-(3-hydroxy-5-tert-butylisoxazol-4-yl) propanoic acid (ATPA) induced significantly larger currents in OXT-mRFP1 neurones than in AVP-eGFP neurones. Furthermore, the difference between the AVP-eGFP and OXT-mRFP1 neurones in the ATPA currents was approximately equal to the difference in the KA currents. These findings suggest that the GluK1-containing KARs may be more highly expressed in OXT neurones than in AVP neurones. These results may provide new insight into the physiology and synaptic plasticity of SON neurones. PMID:24341559

Ohkubo, J; Ohbuchi, T; Yoshimura, M; Maruyama, T; Ishikura, T; Matsuura, T; Suzuki, H; Ueta, Y

2014-01-01

227

Pharmacogenetic heterogeneity of transgene expression in muscle and tumours  

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Abstract Background Recombinant adenoviruses are employed to deliver a therapeutic transgene in the liver, muscle or tumour tissue. However, to rationalise this delivery approach, the factors of variation between individuals need to be identified. It is assumed that differences between inbred strains of laboratory animals are considered to reflect differences between patients. Previously we showed that transgene expression in the liver of different rat strains was dependent o...

Attema Joline; Lefesvre Pierre; van Bekkum Dirk

2003-01-01

228

Generation of Transgenic Mice  

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This unit describes detailed step-by-step protocols, reagents, and equipment required for successful generation of transgenic mice using pronuclear injection. The experimental methods and practical tips given here will help guide beginners in understanding what is required and what to avoid in these standard protocols for efficiently generating transgenic mice.

Cho, Andrew; Haruyama, Naoto; Kulkarni, Ashok B.

2009-01-01

229

WEEDING IN TRANSGENES  

Science.gov (United States)

Transgenes promise to reduce insecticide and fungicide use, but relatively little has been done to significantly reduce herbicide use through genetic engineering. Three strategies for transgene utilization are discussed which have the potential to change this: 1) improvement of weed-specific biocon...

230

Analysis of gene expression changes in relation to toxicity and tumorigenesis in the livers of Big Blue transgenic rats fed comfrey (Symphytum officinale)  

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Abstract Background Comfrey is consumed by humans as a vegetable and a tea, and has been used as an herbal medicine for more than 2000 years. Comfrey, however, is hepatotoxic in livestock and humans and carcinogenic in experimental animals. Our previous study suggested that comfrey induces liver tumors by a genotoxic mechanism and that the pyrrolizidine alkaloids in the plant are responsible for mutation induction and tumor initiation in rat liver. Results In th...

Mei Nan; Guo Lei; Zhang Lu; Shi Leming; Sun Yongming; Fung Chris; Moland Carrie L; Dial Stacey L; Fuscoe James C; Chen Tao

2006-01-01

231

Characterisation of a transgenic mouse expressing R122H human cationic trypsinogen  

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Abstract Background The R122H mutation of the cationic trypsinogen was found in patients with hereditary pancreatitis. A transgenic animal carrying this mutation could be useful as a genetic model system of pancreatitis. Methods Mice transgenic for the human R122H cationic trypsinogen were generated using the -205 fragment of the rat elastase promoter. The presence of the transgene was assayed in the DNA, in pancreatic mRNA and in zymogen granule lysates. Serum ...

Mössner Joachim; Savkovic Vuk; Klöppel Günter; Gaiser Sebastian; Sack Ulrich; Selig Lena; Keim Volker; Bödeker Hans

2006-01-01

232

Tet-Transgenic Rodents: a comprehensive, up-to date database  

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Here we introduce the "Tet-Transgenic Rodents" database, documenting most of the published Tet-transgenic mouse lines generated in the past 2 decades. Aside from the >500 mouse lines listed, it also includes the first of the recently reported Tet-transgenic rat models. Since the Tet technology comprises two essential components, a cis-acting promoter (P(tet)) and a trans-acting transactivator, the database has been organized accordingly. One section of the database summarizes the different tr...

Schoenig, K.; Freundlieb, S.; Gossen, M.

2013-01-01

233

Transgenic mice: beyond the knockout  

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Transgenic mice have had a tremendous impact on biomedical research. Most researchers are familiar with transgenic mice that carry Cre recombinase (Cre) and how they are used to create conditional knockouts. However, some researchers are less familiar with many of the other types of transgenic mice and their applications. For example, transgenic mice can be used to study biochemical and molecular pathways in primary cultures and cell suspensions derived from transgenic mice, cell-cell interac...

Miller, R. Lance

2011-01-01

234

Dual transplantation of human neural stem cells into cervical and lumbar cord ameliorates motor neuron disease in SOD1 transgenic rats  

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Stem cells provide novel sources of cell therapies for motor neuron disease that have recently entered clinical trials. In the present study, we transplanted human neural stem cells (NSCs) into the ventral horn of both the lumbar (L4–L5) and cervical (C4–C5) protuberance of SOD G93A rats, in an effort to test the feasibility and general efficacy of a dual grafting paradigm addressing several muscle groups in the front limbs, hind limbs and the respiratory apparatus. Transplantation was do...

Xu, Leyan; Shen, Peilin; Hazel, Thomas; Johe, Karl; Koliatsos, Vassilis E.

2011-01-01

235

Transgenic crops in Spain.  

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The rate of introduction of transgenic crops in Spanish agriculture has been limited by a number of adverse factors, some of which are linked to specific local circumstances, while others are common to most European Union (EU) member countries. Because of its dry climate, Spain is the main European importer of feed grains, mainly soybeans and corn. The public was introduced to transgenic crops through the appearance of press headlines reporting demonstrations by nongovernmental organizations ...

Garci?a Olmedo, Francisco

2003-01-01

236

Transgenic animal models for the functional analysis of vasoactive peptides  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english The interplay of vasoactive peptide systems is an essential determinant of blood pressure regulation in mammals. While the endothelin and the renin-angiotensin systems raise blood pressure by inducing vasoconstriction and sodium retention, the kallikrein-kinin and the natriuretic-peptide systems red [...] uce arterial pressure by eliciting vasodilatation and natriuresis. Transgenic technology has proven to be very useful for the functional analysis of vasoactive peptide systems. As an outstanding example, transgenic rats overexpressing the mouse Ren-2 renin gene in several tissues become extremely hypertensive. Several other transgenic rat and mouse strains with genetic modifications of components of the renin-angiotensin system have been developed in the past decade. Moreover, in recent years gene-targeting technology was employed to produce mouse strains lacking these proteins. The established animal models as well as the main insights gained by their analysis are summarized in this review.

M., Bader.

1998-09-01

237

Phytoremediation with transgenic trees  

Energy Technology Data Exchange (ETDEWEB)

In the present paper actual trends in the use of transgenic trees for phytoremediation of contaminated soils are reviewed. In this context a current field trial in which transgenic poplars with enhanced GSH synthesis and hence elevated capacity for phytochelatin production are compared with wildtype plants for the removal of heavy metals at different levels of contamination and under different climatic conditions. The studies are carried out with grey poplar (Populus tremula x P. alba), wildtype plants and plants overexpressing the gene for {gamma}-glutamylcysteine synthetase (gshI) from E. coli in the cytosol. The expression of this gene in poplar leads to two- to four-fold enhanced GSH concentrations in the leaves. In greenhouse experiments under controlled conditions these transgenic poplars showed a high potential for uptake and detoxification of heavy metals and pesticides. This capacity is evaluated in field experiments. Further aims of the project are to elucidate (a) the stability of the transgene under field conditions and (b) the possibility of horizontal gene transfer to microorganisms in the rhizosphere. The results will help to assess the biosafety risk of the use of transgenic poplar for phytoremediation of soils. (orig.)

Peuke, A.D.; Rennenberg, H. [Inst. fuer Forstbotanik und Baumphysiologie, Professur fuer Baumphysiologie, Freiburg im Breisgau (Germany)

2005-04-01

238

Deterioration of kidney function by the (pro)renin receptor blocker handle region peptide in aliskiren-treated diabetic transgenic (mRen2)27 rats.  

Science.gov (United States)

Dual renin-angiotensin system (RAS) blockade in diabetic nephropathy is no longer feasible because of the profit/side effect imbalance. (Pro)renin receptor [(P)RR] blockade with handle region peptide (HRP) has been reported to exert beneficial effects in various diabetic models in a RAS-independent manner. To what degree (P)RR blockade adds benefits on top of RAS blockade is still unknown. In the present study, we treated diabetic TGR(mREN2)27 rats, a well-established nephropathy model with high prorenin levels [allowing continuous (P)RR stimulation in vivo], with HRP on top of renin inhibition with aliskiren. Aliskiren alone lowered blood pressure and exerted renoprotective effects, as evidenced by reduced glomerulosclerosis, diuresis, proteinuria, albuminuria, and urinary aldosterone levels as well as diminished renal (P)RR and ANG II type 1 receptor expression. It also suppressed plasma and tissue RAS activity and suppressed cardiac atrial natriuretic peptide and brain natriuretic peptide expression. HRP, when given on top of aliskiren, did not alter the effects of renin inhibition on blood pressure, RAS activity, or aldosterone. However, it counteracted the beneficial effects of aliskiren in the kidney, induced hyperkalemia, and increased plasma plasminogen activator-inhibitor 1, renal cyclooxygenase-2, and cardiac collagen content. All these effects have been linked to (P)RR stimulation, suggesting that HRP might, in fact, act as a partial agonist. Therefore, the use of HRP on top of RAS blockade in diabetic nephropathy is not advisable. PMID:24694588

te Riet, Luuk; van den Heuvel, Mieke; Peutz-Kootstra, Carine J; van Esch, Joep H M; van Veghel, Richard; Garrelds, Ingrid M; Musterd-Bhaggoe, Usha; Bouhuizen, Angelique M; Leijten, Frank P J; Danser, A H Jan; Batenburg, Wendy W

2014-05-15

239

[Visualization of the response in the central nervous system after nociceptive stimulation using transgenic animals].  

Science.gov (United States)

Physiological response to acute and chronic nociceptive stimulation are important for living organisms. In our laboratory, we generated transgenic rats expressing the arginine vasopressin (AVP) and enhanced green fluorescent protein (eGFP) fusion gene, and the c-fos and monomeric red fluorescent protein 1 (mRFP1) fusion gene in the central nervous system. We made it possible to visualize the pain response in the living cells. Using these transgenic rats, the aim of our studies is the elucidation of the physiological role of AVP after nociceptive stimulation and the pathophysiology of work-related pain. We describe the previous findings of nociceptive response, using these transgenic animals. PMID:23270255

Ishikura, Toru; Suzuki, Hitoshi; Matsuura, Takanori; Ohnishi, Hideo; Nakamura, Toshitaka; Ueta, Yoichi

2012-12-01

240

Transgenic mice susceptible to poliovirus.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Poliovirus-sensitive transgenic mice were produced by introducing the human gene encoding cellular receptors for poliovirus into the mouse genome. Expression of the receptor mRNAs in tissues of the transgenic mice was analyzed by using RNA blot hybridization and the polymerase chain reaction. The human gene is expressed in many tissues of the transgenic mice just as in tissues of humans. The transgenic mice are susceptible to all three poliovirus serotypes, and the mice inoculated with poliov...

Koike, S.; Taya, C.; Kurata, T.; Abe, S.; Ise, I.; Yonekawa, H.; Nomoto, A.

1991-01-01

 
 
 
 
241

Transgenic Crops for Herbicide Resistance  

Science.gov (United States)

Since their introduction in 1995, crops made resistant to the broad-spectrum herbicides glyphosate and glufosinate with transgenes are widely available and used in much of the world. As of 2008, over 80% of the transgenic crops grown world-wide have this transgenic trait. This technology has had m...

242

Calcium electrotransfer for termination of transgene expression in muscle.  

Science.gov (United States)

Gene electrotransfer is expanding in clinical use, thus we have searched for an emergency procedure to stop transgene expression in case of serious adverse events. Calcium is cytotoxic at high intracellular levels, so we tested effects of calcium electrotransfer on transgene expression in muscle. A clinical grade calcium solution (20 ?l, 168 mM) was injected into transfected mouse or rat tibialis cranialis muscle. Ca(2+) uptake was quantified using calcium 45 ((45)Ca), and voltage and time between injection and pulsation were varied. Extinction of transgene expression was investigated by using both in vivo imaging of infrared fluorescent "Katushka" and erythropoietin evaluated by ELISA and hemoglobin. Histology was performed. Electrotransfer of Katushka and erythropoietin yielded significant expression. Maximal calcium uptake occurred after injection of Ca(2+) before electropulsing using eight high voltage pulses of 1000 V/cm. Using these parameters, in vivo imaging showed that transgene expression significantly decreased 4 hr after Ca(2+) electrotransfer and was eliminated within 24 hr. Similarly, serum erythropoietin was reduced by 46% at 4 hr and to control levels at 2 days. Histological analyses showed muscle damage and subsequent regeneration. Electrotransfer of isotonic CaCl(2) terminates transgenic protein expression in muscles and may be used for contingency elimination of transgene expression. PMID:21470044

Hojman, Pernille; Spanggaard, Iben; Olsen, Caroline Holkman; Gehl, Julie; Gissel, Hanne

2011-06-01

243

Calcium electrotransfer for termination of transgene expression in muscle  

DEFF Research Database (Denmark)

Gene electrotransfer is expanding in clinical use, thus we have searched for an emergency procedure to stop transgene expression in case of serious adverse events. Calcium is cytotoxic at high intracellular levels, so we tested effects of calcium electrotransfer on transgene expression in muscle. A clinical grade calcium solution (20 ?l, 168 mM) was injected into transfected mouse or rat tibialis cranialis muscle. Ca(2+) uptake was quantified using calcium 45 ((45)Ca), and voltage and time between injection and pulsation were varied. Extinction of transgene expression was investigated by using both in vivo imaging of infrared fluorescent "Katushka" and erythropoietin evaluated by ELISA and hemoglobin. Histology was performed. Electrotransfer of Katushka and erythropoietin yielded significant expression. Maximal calcium uptake occurred after injection of Ca(2+) before electropulsing using eight high voltage pulses of 1000 V/cm. Using these parameters, in vivo imaging showed that transgene expression significantly decreased 4 hr after Ca(2+) electrotransfer and was eliminated within 24 hr. Similarly, serum erythropoietin was reduced by 46% at 4 hr and to control levels at 2 days. Histological analyses showed muscle damage and subsequent regeneration. Electrotransfer of isotonic CaCl(2) terminates transgenic protein expression in muscles and may be used for contingency elimination of transgene expression.

Hojman, Pernille; Spanggaard, Iben

2011-01-01

244

Generation of transgenic frogs.  

Science.gov (United States)

The possibility of generating transgenic animals is of obvious advantage for the analysis of gene function in development and disease. One of the established vertebrate model systems in developmental biology is the amphibian Xenopus laevis. Different techniques have been successfully applied to create Xenopus transgenics; in this chapter, the so-called meganuclease method is described. This technique is not only technically simple, but also comparably efficient and applicable to both Xenopus laevis and Xenopus tropicalis. The commercially available endonuclease I-SceI (meganuclease) mediates the integration of foreign DNA into the frog genome after coinjection into fertilized eggs. Tissue-specific gene expression, as well as germline transmission, has been observed. PMID:19504064

Loeber, Jana; Pan, Fong Cheng; Pieler, Tomas

2009-01-01

245

Retinoblastoma in transgenic mice  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Retinoblastoma, a malignancy of the eye occurring in young children, has been widely studied as a model for genetic predisposition to cancer. This disease is caused by mutations in both alleles of an anti-oncogene (the retinoblastoma gene, Rb) that inactivate or eliminate the Rb encoded protein, pl05rb. Here we report that expression of a viral oncogene, the simian virus 40 T antigen, in the retina of transgenic mice produces heritable ocular tumours with histological, ul...

Windle, J. J.; Albert, D. M.; O Brien, J. M.; Marcus, D. M.; Disteche, Ch M.; Bernards, R. A.; Mellon, P. L.

1990-01-01

246

Transgenics in crops  

Science.gov (United States)

With rapid world population growth and declining availability of fresh water and arable land, a new technology is urgently needed to enhance agricultural productivity. Recent discoveries in the field of crop transgenics clearly demonstrate the great potential of this technology for increasing food production and improving food quality while preserving the environment for future generations. In this review, we briefly discuss some of the recent achievements in crop improvement that have been made using gene transfer technology.

Li, Y.; Wu, Y. H.; McAvoy, R.; Duan, H.

2001-01-01

247

COMPARISON TRANSGENIC AND NON-TRANSGENIC MILK QUALITY  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Transgenic founder rabbits carrying a gene construct consisting of a 2.5 kb murine whey acidic protein promoter (mWAP), 7.2 kb of the human clotting factor VIII (hFVIII) cDNA and 4.6 kb of 3’ flanking sequences of mWAP gene were crossed for five generations. Transgenic females showed high level of recombinant hFVIII (rhFVIII) mRNA expression in biopsed mammary gland tissues. The presence of the mWAP-hFVIII transgene in rabbit genome and secretion of rhFVIII into milk of transgenic females (...

Peter Chrenek; Alexander Makarevich2

2012-01-01

248

Transgenic algae engineered for higher performance  

Energy Technology Data Exchange (ETDEWEB)

The present disclosure relates to transgenic algae having increased growth characteristics, and methods of increasing growth characteristics of algae. In particular, the disclosure relates to transgenic algae comprising a glutamine phenylpyruvate transaminase transgene and to transgenic algae comprising a glutamine phenylpyruvate transaminase transgene and a glutamine synthetase.

Unkefer, Pat J; Anderson, Penelope S; Knight, Thomas J

2014-10-21

249

Transgenic mice expressing beta-galactosidase in mature neurons under neuron-specific enolase promoter control.  

Science.gov (United States)

To gain insights into transcription factors defining neuronal identity, we generated transgenic mice carrying a 1.8 kb rat neuron-specific enolase (NSE) promoter fragment fused to an E. coli lacZ gene. Four of seven transgenic families expressed transgene RNA in the nervous system but not in most other tissues. Histochemical analysis of adult brain from the two lines with highest lacZ mRNA levels showed neuron-specific, pan-neuronal beta-galactosidase activity. Developmental RNA and histochemical analyses showed parallel onset of transgene and endogenous NSE gene expression in various neuronal cell types, although the magnitude of NSE mRNA accumulation later in development was not matched by the transgene. These results suggest that cis-acting regulatory elements, subject to neuron-specific control, are located within 1.8 kb upstream from the NSE gene. PMID:2116814

Forss-Petter, S; Danielson, P E; Catsicas, S; Battenberg, E; Price, J; Nerenberg, M; Sutcliffe, J G

1990-08-01

250

L-selectin can facilitate metastasis to lymph nodes in a transgenic mouse model of carcinogenesis  

Digital Repository Infrastructure Vision for European Research (DRIVER)

L-selectin mediates homing of lymphocytes to lymph nodes (LN). Transgenic mice that express rat insulin promoter regulated simian virus 40 Tag (RIP-Tag) develop large, local cancers that metastasize to liver but not LN. To test whether this lack of LN metastases reflects their absence from the circulation, transgenic mice were produced that express Tag (T), L-selectin (L), and Escherichia coli LacZ (Z), in pancreatic ? cells. LTZ mice developed insulinomas that sp...

Qian, Fawn; Hanahan, Douglas; Weissman, Irving L.

2001-01-01

251

Transgenic A1 adenosine receptor overexpression increases myocardial resistance to?ischemia  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Activation of myocardial A1 adenosine receptors (A1AR) protects the heart from ischemic injury. In this study transgenic mice were created using the cardiac-specific ?-myosin heavy chain promoter and rat A1AR cDNA. Heart membranes from two transgene positive lines displayed ?1,000-fold overexpression of A1AR (6,574 ± 965 and 10,691 ± 1,002 fmol per mg of protein vs. 8 ± 5 fmol per mg of protein in control hearts). Compared with control hearts, transgenic Langendorff-perfused hearts had ...

Matherne, G. Paul; Linden, Joel; Byford, Anne M.; Gauthier, Naomi S.; Headrick, John P.

1997-01-01

252

Plasma lipoprotein metabolism in transgenic mice overexpressing apolipoprotein E. Accelerated clearance of lipoproteins containing apolipoprotein B.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

We have reported that transgenic mice overexpressing rat apo E shows marked reduction of plasma cholesterol and triglyceride levels due to the disappearance of VLDL and LDL. In this study, we investigated the metabolism of plasma lipoproteins in transgenic mice. After intravenous injection, the rates of clearance of 125I-VLDL and 125I-LDL were 3.0- and 2.4-fold greater in transgenic mice than in controls, respectively. Furthermore, clearance of chylomicron remnants estimated by oral retinyl p...

Shimano, H.; Yamada, N.; Katsuki, M.; Yamamoto, K.; Gotoda, T.; Harada, K.; Shimada, M.; Yazaki, Y.

1992-01-01

253

TL transgenic mouse strains  

International Nuclear Information System (INIS)

As a result of abnormal development of the thymus of these mice, TCR ?? lineage of the T cell differentiation is disturbed and cells belonging to the TCR ?? CD4- CD8- double negative (DN) lineage become preponderant. The ?? DN cells migrate into peripheral lymphoid organs and constitute nearly 50% of peripheral T cells. Immune function of the transgenic mice is severely impaired, indicating that the ?? cells are incapable of participating in these reactions. Molecular and serological analyses of T-cell lymphomas reveal that they belong to the ?? lineage. Tg.Tlaa-3-1 mice should be useful in defining the role of TL in normal and abnormal T cell differentiation as well as in the development of T-cell lymphomas, and further they should facilitate studies on the differentiation and function of ?? T cells. We isolated T3b-TL gene from B6 mice and constructed a chimeric gene in which T3b-TL is driven by the promoter of H-2Kb. With the chimeric gene, two transgenic mouse strains, Tg. Con.3-1 and -2 have been derived in C3H background. Both strains express TL antigen in various tissues including skin. The skin graft of transgenic mice on C3H and (B6 X C3H)F1 mice were rejected. In the mice which rejected the grafts, CD8+TCR?? cytotoxic T cells (CTL) against TL antigens were recognized. The recognition of TL by CTL did not require the antigen presentation by H-2 molecules. Thetation by H-2 molecules. The results indicated that TL antigen in the skin becomes a transplantation antigen and behaves like a typical allogeneic MHC class I antigen. The facts that (B6 X C3H)F1 mice rejected the skin expressing T3b-TL antigen and induced CTL that killed TL+ lymphomas of B6 origin revealed that TL antigen encoded by T3b-TL is recognized as non-self in B6 mice. Experiments are now extended to analyze immune responses to TL antigen expressed on autochthonous T cell lymphomas. (J.P.N.)

254

Improved Cre reporter transgenic Xenopus  

Digital Repository Infrastructure Vision for European Research (DRIVER)

We have produced and characterized improved transgenic reporter lines for detection of Cre recombinase activity during Xenopus development. Improvements include choice of fluorophores, which make these Cre reporter lines generally suitable for lineage tracing studies. We also include data for several new parameters affecting survival and transgenesis efficiency using the recently developed meganuclease method of frog transgenesis. These transgenic frogs express cyan fluorescent protein (CFP) ...

Rankin, Scott A.; Hasebe, Takashi; Zorn, Aaron M.; Buchholz, Daniel R.

2009-01-01

255

A qRT-PCR RFLP Method for Monitoring Highly Conserved Transgene Expression during Gene Therapy  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Evaluation of the transfer efficiency of a rat heme oxygenase-1 (HO-1) transgene into mice requires differentiation of rat and mouse HO-1. However, rat and mouse HO-1 have 94% homology; antibodies and enzyme activity cannot adequately distinguish HO-1. We designed a qRT-PCR method to monitor HO-1 transcription relative to a housekeeping gene, GAPDH. The ratio of rat and mouse HO-1 mRNA could be estimated through restriction fragment length polymorphism (RFLP) analysis of the PCR products. In ...

Bruzzone, Carol M.; Belcher, John D.; Schuld, Nathan J.; Newman, Kristal A.; Vineyard, Julie; Nguyen, Julia; Chen, Chunsheng; Beckman, Joan D.; Steer, Clifford J.; Vercellotti, Gregory M.

2008-01-01

256

ALIMENTOS TRANSGÉNICOS TRANSGENIC FOODS  

Directory of Open Access Journals (Sweden)

Full Text Available Gracias al gran avance de la tecnología, la ingeniería genética y la biología molecular, se han desarrollado los productos transgénicos. En sus inicios, los productos modificados genéticamente tenían como objeto obtener ventajas en las áreas de la agricultura y ganadería. Posteriormente esta técnica se comenzó a aplicar en el ámbito de la producción de alimentos para el consumo humano. Se ha generado mucha controversia en relación a su utilización. Esta revisión tiene por objeto revisar la información científica disponible en relación a las aplicaciones, ventajas y potenciales riesgos para la salud humana y el medio ambiente asociados al consumo de los alimentos transgénicosDue to the advancements in technology, genetic engineering and molecular biology, have develop transgenic foods. Initially, genetically modified plants were produced to confer advantages in agriculture and animal husbandry. Later this technique was applied to the production of food for human consumption, generating a great deal of controversy. This review discusses the available scientific evidence in relation to the advantages and potential risks of genetically modified foods

María Soledad Reyes S.

2003-04-01

257

ALIMENTOS TRANSGÉNICOS / TRANSGENIC FOODS  

Scientific Electronic Library Online (English)

Full Text Available SciELO Chile | Language: Spanish Abstract in spanish Gracias al gran avance de la tecnología, la ingeniería genética y la biología molecular, se han desarrollado los productos transgénicos. En sus inicios, los productos modificados genéticamente tenían como objeto obtener ventajas en las áreas de la agricultura y ganadería. Posteriormente esta técnica [...] se comenzó a aplicar en el ámbito de la producción de alimentos para el consumo humano. Se ha generado mucha controversia en relación a su utilización. Esta revisión tiene por objeto revisar la información científica disponible en relación a las aplicaciones, ventajas y potenciales riesgos para la salud humana y el medio ambiente asociados al consumo de los alimentos transgénicos Abstract in english Due to the advancements in technology, genetic engineering and molecular biology, have develop transgenic foods. Initially, genetically modified plants were produced to confer advantages in agriculture and animal husbandry. Later this technique was applied to the production of food for human consump [...] tion, generating a great deal of controversy. This review discusses the available scientific evidence in relation to the advantages and potential risks of genetically modified foods

María Soledad, Reyes S.; Jaime, Rozowski N.

2003-04-01

258

Intracerebral transplants of primary muscle cells: a potential 'platform' for transgene expression in the brain  

Science.gov (United States)

After the transplantation of rat primary muscle cells into the caudate or cortex of recipient rats, the muscle cells were able to persist for at least 6 months. Muscle cells transfected with expression plasmids prior to transplantation were able to express reporter genes in the brains for at least 2 months. These results suggest that muscle cells might be a useful 'platform' for transgene expression in the brain.

Jiao, S.; Schultz, E.; Wolff, J. A.

1992-01-01

259

Derivation of a Germline Competent Transgenic Fischer 344 Embryonic Stem Cell Line  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Embryonic stem (ES) cell-based gene manipulation is an effective method for the generation of mutant animal models in mice and rats. Availability of germline-competent ES cell lines from inbred rat strains would allow for creation of new genetically modified models in the desired genetic background. Fischer344 (F344) males carrying an enhanced green fluorescence protein (EGFP) transgene were used as the founder animals for the derivation of ES cell lines. After establishment of ES cell lines,...

Men, Hongsheng; Bryda, Elizabeth C.

2013-01-01

260

Cloning of the goat beta-casein-encoding gene and expression in transgenic mice.  

Science.gov (United States)

The goat beta-casein-encoding gene (CSN2), which encodes the most abundant protein of goat milk, has been cloned and sequenced. The intron/exon organization of the 9.0-kb goat CSN2 gene is similar to that of other CSN2 genes. Expression of the goat gene was principally restricted to the mammary gland of lactating transgenic animals. A low level of expression was also observed in skeletal muscle and skin. In contrast to a rat CSN2 transgene [Lee et al., Nucleic Acids Res. 16 (1988) 1027-1041], the goat gene was expressed to a high degree in the lactating mammary gland. Differences in the content or context of regulatory elements may account for the enhanced performance of the goat relative to the rat CSN2 gene in transgenic mice. PMID:1446822

Roberts, B; DiTullio, P; Vitale, J; Hehir, K; Gordon, K

1992-11-16

 
 
 
 
261

Temporal Expression of Mutant LRRK2 in Adult Rats Impairs Dopamine Reuptake  

Directory of Open Access Journals (Sweden)

Full Text Available Parkinson's disease (PD results from progressive degeneration of dopaminergic neurons. Most PD cases are sporadic, but some have pathogenic mutation in the individual genes. Mutation of the leucine-rich repeat kinase-2 (LRRK2 gene is associated with familial and sporadic PD, as exemplified by G2019S substitution. While constitutive expression of mutant LRRK2 in transgenic mice fails to induce neuron death, transient expression of the disease gene by viral delivery causes a substantial loss of dopaminergic neurons in mice. To further assess LRRK2 pathogenesis, we created inducible transgenic rats expressing human LRRK2 with G2019S substitution. Temporal overexpression of LRRK2G2019S in adult rats impaired dopamine reuptake by dopamine transporter (DAT and thus enhanced locomotor activity, the phenotypes that were not observed in transgenic rats constitutively expressing the gene throughout life time. Reduced DAT binding activity is an early sign of dopaminergic dysfunction in asymptomatic subjects carrying pathogenic mutation in LRRK2. Our transgenic rats recapitulated the initiation process of dopaminergic dysfunction caused by pathogenic mutation in LRRK2. Inducible transgenic approach uncovered phenotypes that may be obscured by developmental compensation in constitutive transgenic rats. Finding in inducible LRRK2 transgenic rats would guide developing effective strategy in transgenic studies: Inducible expression of transgene may induce greater phenotypes than constitutive gene expression, particularly in rodents with short life time.

Hongxia Zhou, Cao Huang, Jianbin Tong, Weimin C Hong, Yong-Jian Liu, Xu-Gang Xia

2011-01-01

262

Pathologic progression of mammary carcinomas in a C3(1)/SV40 T/t-antigen transgenic rat model of human triple-negative and Her2-positive breast cancer  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The C3(1) component of the rat prostate steroid binding protein has been used to target expression of the SV40 T/t-antigen to the mammary epithelium of mice resulting in pre-neoplastic lesions that progress to invasive and metastatic cancer with molecular features of human basal-type breast cancer. However, there are major differences in the histologic architecture of the stromal and epithelial elements between the mouse and human mammary glands. The rat mammary gland is more enriched with ep...

Hoenerhoff, M. J.; Shibata, M. A.; Bode, A.; Green, J. E.

2011-01-01

263

Transgenic RNA Interference in Mice  

Science.gov (United States)

The discovery that small interfering RNA duplexes (siRNA) can silence gene expression in mammalian cells has revolutionized biomedical research. The most successful application of the discovery has been to study gene function in cultured human or mouse cells. However, the knockdown effect of siRNA is only transient. To achieve a more sustained gene-silencing effect, shRNA (small hairpin RNA) expressed from a vector is preferred. An additional benefit of shRNA is that RNA interference (RNAi) can now be applied in vivo through delivering shRNA-expressing vectors by transgenic technology. Transgenic RNAi not only allows the study of biological processes not present in cultured cells but also offers chronic therapeutic potentials. In this review, we will summarize the developments in the generation of transgenic RNAi mice.

2007-06-01

264

Transgenic agriculture and environmental indicators  

Directory of Open Access Journals (Sweden)

Full Text Available Despite the rapid diffusion of transgenic crops, there are still few environmental impact studies capable of supplying a conclusive scientific response in regard to its technical and economic advantages and disadvantages. Prospective scenarios were elaborated to assist environmental impact assessment, using techniques derived from SWOT (Strength, Weakness, Opportunity, Threat analysis and the DPSIR (Driving Force – human activity, Pressure, State, Impact, Response model, to evaluate the environmental indicators and the relationship between them. Control and management actions were identified, searching the integration of aspects related to the biotechnology applied to transgenic processes, biodiversity, biosafety and intellectual property. It was demonstrated that the DPSIR model is, in fact, an instrument for integrated environmental assessment and the application of the proposed methodology resulted in favorable indicators to the adoption of transgenic agriculture. The elaborated scenarios are useful to develop an Environmental Management System (EMS to agriculture.

Denize Dias de Carvalho

2006-12-01

265

How To Produce and Characterize Transgenic Plants.  

Science.gov (United States)

Explains the process of establishing transgenic plants which is a very important tool in plant biology and modern agriculture. Produces transgenic plants with the ability to synthesize opines. (Contains 17 references.) (YDS)

Savka, Michael A.; Wang, Shu-Yi; Wilson, Mark

2002-01-01

266

Transgenic crops, production risk, and agrobiodiversity  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Do transgenic crops cause agrobiodiversity erosion? We hypothesize that they increase productivity and reduce production risk and may therefore reduce farmer demand for on-farm varietal diversity, especially when only a few transgenic varieties are available. We also hypothesize that varietal diversity can be preserved when more transgenic varieties are supplied. These hypotheses are tested and confirmed with panel data for the case of transgenic cotton in India. Cotton varietal diversity in ...

Krishna Vijesh; Qaim, Matin; Zilberman, David

2014-01-01

267

Transgenic overexpression of connexin50 induces cataracts  

Digital Repository Infrastructure Vision for European Research (DRIVER)

To examine the effects of increased expression of Cx50 in the mouse lens, transgenic mice were generated using a DNA construct containing the human Cx50 coding region and a C-terminal FLAG epitope driven by the chicken ?B1-crystallin promoter. Expression of this protein in paired Xenopus oocytes induced gap junctional currents of similar magnitude to wild type human Cx50. Three lines of transgenic mice expressing the transgenic protein were analyzed. Lenses from transgenic mice were smaller ...

Chung, June; Berthoud, Viviana M.; Novak, Layne; Zoltoski, Rebecca; Heilbrunn, Benjamin; Minogue, Peter J.; Liu, Xiaoqin; Ebihara, Lisa; Kuszak, Jer; Beyer, Eric C.

2007-01-01

268

HLA-B27 and gender independently determine the likelihood of a positive MRI of the sacroiliac joints in patients with early inflammatory back pain : a 2-year MRI follow-up study  

DEFF Research Database (Denmark)

To describe how inflammation on MRI of the sacroiliac joints in patients with recent-onset inflammatory back pain (IBP) evolves over time, and to study determinants of activity on MRI of the sacroiliac joint.

van Onna, M; Jurik, A G

2011-01-01

269

Progress on researches of transgenic alfalfa  

International Nuclear Information System (INIS)

In this paper, the progress on the researches of transgenic alfalfa in the past two decades had been reviewed in the aspects of regeneration system, transformation, improvement of the important traits and so on. Moreover, such problems as variation of transgene expression and safety of transgenic plant had also been discussed and propose had been given for the future research work. (authors)

270

Variegated transgene expression in mouse mammary gland is determined by the transgene integration locus.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Mice carrying an ovine beta-lactoglobulin (BLG) transgene secrete BLG protein into their milk. To explore transgene expression stability, we studied expression levels in three BLG transgenic mouse lines. Unexpectedly, two lines exhibited variable levels of transgene expression. Copy number within lines appeared to be stable and there was no evidence of transgene rearrangement. In the most variable line, BLG production levels were stable within individual mice in two successive lactations. Bac...

Dobie, K. W.; Lee, M.; Fantes, J. A.; Graham, E.; Clark, A. J.; Springbett, A.; Lathe, R.; Mcclenaghan, M.

1996-01-01

271

Age and lesion-induced increases of GDNF transgene expression in brain following intracerebral injections of DNA nanoparticles.  

Science.gov (United States)

In previous studies that used compacted DNA nanoparticles (DNP) to transfect cells in the brain, we observed higher transgene expression in the denervated striatum when compared to transgene expression in the intact striatum. We also observed that long-term transgene expression occurred in astrocytes as well as neurons. Based on these findings, we hypothesized that the higher transgene expression observed in the denervated striatum may be a function of increased gliosis. Several aging studies have also reported an increase of gliosis as a function of normal aging. In this study we used DNPs that encoded for human glial cell line-derived neurotrophic factor (hGDNF) and either a non-specific human polyubiquitin C (UbC) or an astrocyte-specific human glial fibrillary acidic protein (GFAP) promoter. The DNPs were injected intracerebrally into the denervated or intact striatum of young, middle-aged or aged rats, and glial cell line-derived neurotrophic factor (GDNF) transgene expression was subsequently quantified in brain tissue samples. The results of our studies confirmed our earlier finding that transgene expression was higher in the denervated striatum when compared to intact striatum for DNPs incorporating either promoter. In addition, we observed significantly higher transgene expression in the denervated striatum of old rats when compared to young rats following injections of both types of DNPs. Stereological analysis of GFAP(+) cells in the striatum confirmed an increase of GFAP(+) cells in the denervated striatum when compared to the intact striatum and also an age-related increase; importantly, increases in GFAP(+) cells closely matched the increases in GDNF transgene levels. Thus neurodegeneration and aging may lay a foundation that is actually beneficial for this particular type of gene therapy while other gene therapy techniques that target neurons are actually targeting cells that are decreasing as the disease progresses. PMID:25453772

Yurek, D M; Hasselrot, U; Cass, W A; Sesenoglu-Laird, O; Padegimas, L; Cooper, M J

2015-01-22

272

Induction of Proteinuria by Cannabinoid Receptors 1 Signaling Activation in CB1 Transgenic Mice.  

Science.gov (United States)

: Proteinuria is not only a sign of kidney damage but is also involved in the progression of renal disease as an independent pathologic factor. Although patients with mutated type 1 cannabinoid receptors (CB1) polymorphism are associated with renal microvascular damage, the biologic role of CB1 signaling in proteinuria remains uncharacterized till now. Herein, we investigate whether CB1 participates in glomerular proteinuria in CB1 transgenic mice and treatment with CB1 agonist WIN55212-2 rat, neither of which are diabetic models. The CB1 transgenic mice and rats treated with CB1 agonist WIN55212-2 had higher kidney weight and urinary protein concentrations but not blood glucose levels compared with the wild-type group. A combination of laser-capture microsdissection, quantitative reverse transcription-polymerase chain reaction, immunoblotting and immunohistochemical validation revealed that CB1 transgenic mice and rats treated with CB1 agonist WIN55212-2 had higher vascular endothelial growth factor (VEGF) expression in renal glomeruli than that of the wild-type group. Geneticorpharmacological activation of CB1 by transgenic CB1 mice or treatment with WIN55212-2 reduced nephrin expression in the renal glomeruli compared with that of the wild-type group in the glomerular mesanglium. Taken together, CB1 transgenic mice and rats treated with CB1 agonist WIN55212-2 induced proteinuria with upregulation of CB1 resulting in impaired nephrin expression, by inducing excess VEGF reaction in the renal glomeruli. Genetic and pharmacological manipulation of CB1 signaling revealed VEGF-dependent nephrin depression of glomerulopathy. Controlling CB1 activity can be used an alternative strategy for sustaining renal function in the presence of CB1 activation. PMID:25474224

Hsu, Yung-Chien; Lei, Chen-Chou; Shih, Ya-Hsueh; Ho, Cheng; Lin, Chun-Liang

2014-12-01

273

[Impact of transgenes and cloning on xenografts].  

Science.gov (United States)

Transgenesis can theoretically add a foreign gene or specifically replace an endogenous gene by another gene. Gene addition in mammals is generally achieved by DNA microinjection into one-cell embryos. Gene replacement implies homologous recombination in cultured cells which must be selected and remain capable of generating a living organism. The use of totipotent cells can, though currently in the mouse only, lead to the generation of chimeric animals transmitting the genetic modification to offsprings. The embryo cloning technique has recently allowed the use of somatic fetal cells, in which gene replacement occurred to generate living sheeps. This technique is being extended to other domestic ruminants, to pigs and rabbits. The mouse, rat and rabbit are being used as models to define the genes which should be added or inactivated to reduce rejections of xenografts. Transgenic pigs harbouring the human CD59 or the DAF genes have been obtained by several groups. Heart, kidney and isolated cells from these animals are more resistant to hyperacute rejection when grafted to experimental primates. Additional genes are to be added in the future to inhibit the other rejection mechanisms as soon as their action has been demonstrated in laboratory animals. Experiments in progress aim at inactivating the gene for Gal: 1-3-galactosyl transferase by homologous recombination in the pig genome. Transgenesis might also be used to prevent expression of endogenous pig retroviral vectors and to prepare recombinant proteins having antirejection activity from the milk of animals. PMID:10868403

Houdebine, L M

2000-05-01

274

Transgenic animals modelling polyamine metabolism-related diseases.  

Science.gov (United States)

Cloning of genes related to polyamine metabolism has enabled the generation of genetically modified mice and rats overproducing or devoid of proteins encoded by these genes. Our first transgenic mice overexpressing ODC (ornithine decarboxylase) were generated in 1991 and, thereafter, most genes involved in polyamine metabolism have been used for overproduction of the respective proteins, either ubiquitously or in a tissue-specific fashion in transgenic animals. Phenotypic characterization of these animals has revealed a multitude of changes, many of which could not have been predicted based on the previous knowledge of the polyamine requirements and functions. Animals that overexpress the genes encoding the inducible key enzymes of biosynthesis and catabolism, ODC and SSAT (spermidine/spermine N1-acetyltransferase) respectively, appear to possess the most pleiotropic phenotypes. Mice overexpressing ODC have particularly been used as cancer research models. Transgenic mice and rats with enhanced polyamine catabolism have revealed an association of rapidly depleted polyamine pools and accelerated metabolic cycle with development of acute pancreatitis and a fatless phenotype respectively. The latter phenotype with improved glucose tolerance and insulin sensitivity is useful in uncovering the mechanisms that lead to the opposite phenotype in humans, Type 2 diabetes. Disruption of the ODC or AdoMetDC [AdoMet (S-adenosylmethionine) decarboxylase] gene is not compatible with mouse embryogenesis, whereas mice with a disrupted SSAT gene are viable and show no harmful phenotypic changes, except insulin resistance at a late age. Ultimately, the mice with genetically altered polyamine metabolism can be used to develop targeted means to treat human disease conditions that they relevantly model. PMID:20095974

Alhonen, Leena; Uimari, Anne; Pietilä, Marko; Hyvönen, Mervi T; Pirinen, Eija; Keinänen, Tuomo A

2009-01-01

275

Transgenic models of Huntington's disease.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Huntington's disease (HD) is an inherited neurodegenerative disorder caused by a CAG-polyglutamine repeat expansion. A mouse model of this disease has been generated by the introduction of exon 1 of the human HD gene carrying highly expanded CAG repeats into the mouse germ line (R6 lines). Transgenic mice develop a progressive neurological phenotype with a movement disorder and weight loss similar to that in HD. We have previously identified neuronal inclusions in the brains of these mice tha...

Sathasivam, K.; Hobbs, C.; Mangiarini, L.; Mahal, A.; Turmaine, M.; Doherty, P.; Davies, S. W.; Bates, G. P.

1999-01-01

276

Transgenic parasites accelerate drug discovery  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Parasitic neglected diseases are in dire need of new drugs either to replace old drugs rendered ineffective because of resistance development, to cover clinical needs that had never been addressed or to tackle other associated problems of existing drugs such as high cost, difficult administration, restricted coverage or toxicity. The availability of transgenic parasites expressing reporter genes facilitates the discovery of new drugs through high throughput screenings, but also by allowing ra...

Rodriguez, Ana; Tarleton, Rick L.

2012-01-01

277

Transgenic Zebrafish Using Transposable Elements  

Digital Repository Infrastructure Vision for European Research (DRIVER)

DNA transposons are effective chromosomal engineering vehicles for making transgenic zebrafish. We describe both autonomous and non-autonomous transposable elements, and we compare and contrast popular transposon systems. The Tol2 system is a robust gene transfer tool and has been selected as the primary transposon platform, facilitating the development of an array of reagents readily shared within the zebrafish community. We present common transposon and transposase vectors within the field ...

Clark, Karl J.; Urban, Mark D.; Skuster, Kimberly J.; Ekker, Stephen C.

2011-01-01

278

Transgenic agriculture and environmental indicators  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Despite the rapid diffusion of transgenic crops, there are still few environmental impact studies capable of supplying a conclusive scientific response in regard to its technical and economic advantages and disadvantages. Prospective scenarios were elaborated to assist environmental impact assessment, using techniques derived from SWOT (Strength, Weakness, Opportunity, Threat) analysis and the DPSIR (Driving Force – human activity, Pressure, State, Impact, Response) model, to evaluate the e...

Denize Dias de Carvalho; Lucila Teresa de Gusmão Pessôa; Nei Pereira Jr.

2006-01-01

279

Transgenic Arabidopsis Gene Expression System  

Science.gov (United States)

The Transgenic Arabidopsis Gene Expression System (TAGES) investigation is one in a pair of investigations that use the Advanced Biological Research System (ABRS) facility. TAGES uses Arabidopsis thaliana, thale cress, with sensor promoter-reporter gene constructs that render the plants as biomonitors (an organism used to determine the quality of the surrounding environment) of their environment using real-time nondestructive Green Fluorescent Protein (GFP) imagery and traditional postflight analyses.

Ferl, Robert; Paul, Anna-Lisa

2009-01-01

280

Malignant melanoma in transgenic mice.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Ocular and cutaneous melanomas arose in new inbred lines of transgenic mice having an integrated recombinant gene comprised of the tyrosinase promoter, expressed in pigment cells, and the simian virus 40 early-region transforming sequences. The tumors were hypomelanotic and were histopathologically similar to corresponding human melanomas. Eye melanomas often originated at a young age, chiefly from the retinal pigment epithelium, also from the choroid, and rarely from the ciliary body. The ey...

Bradl, M.; Klein-szanto, A.; Porter, S.; Mintz, B.

1991-01-01

 
 
 
 
281

NINDS GENSAT BAC Transgenic Project  

Science.gov (United States)

This website from Rockefeller University in New York contains "a gene expression atlas of the central nervous system of the mouse based on bacterial artificial chromosomes (BACs)." GENSAT, or the Gene Expression Nervous System Atlas, contains brain slice images of BAC transgenic mice at the embryonic, postnatal (7 days old), and adult stages, stained to show areas of gene activity. The website comes with a detailed and helpful tutorial that recreates GENSAT's user interface and demonstrates how to manipulate search results.

282

Improved cre reporter transgenic Xenopus.  

Science.gov (United States)

We have produced and characterized improved transgenic reporter lines for detection of Cre recombinase activity during Xenopus development. Improvements include choice of fluorophores, which make these Cre reporter lines generally suitable for lineage tracing studies. We also include data for several new parameters affecting survival and transgenesis efficiency using the recently developed meganuclease method of frog transgenesis. These transgenic frogs express cyan fluorescent protein (CFP) under control of the ubiquitous promoter CMV, where CFP is replaced by DsRed2 (a red fluorescent protein) in the presence of Cre. Three independent, high expression, Cre-sensitive lines have been identified that maintain robust fluorophore expression across generations and lack DsRed2 expression in the absence of Cre. A novel use of these lines is to indelibly mark embryonic blastomeres by Cre mRNA injection for permanent fate mapping. Similarly, transgenically expressed Cre under control of tissue-specific promoters will allow detailed analysis of cell lineage relationships throughout embryogenesis, metamorphosis, and adulthood. PMID:19653309

Rankin, Scott A; Hasebe, Takashi; Zorn, Aaron M; Buchholz, Daniel R

2009-09-01

283

Transgenic trees and forestry biosafety  

Scientific Electronic Library Online (English)

Full Text Available SciELO Chile | Language: English Abstract in english The benefits from the development of transgenic trees are expected from the improvement of traits as growth and form, wood quality, industrial processes, disease and insect resistance, herbicide tolerance, ecological restoration, rooting ability, etc. One of the first reported field trials with gene [...] tically modified forest trees was established in Belgium in 1988 and the characteristic evaluated was herbicide tolerance in poplars. Since then, there have been more than 200 reported trials, involving at least 15 forest species. The majority of the field trials have been carried out in the USA (64%). More than 50% of the field trials are done with Populus species and the main target traits are herbicide tolerance (31%), followed by marker genes (23%) and insect resistance (14%). Until today, there is only one report on commercial-scale production of transgenic forest trees which is Populus nigra with the Bt gene release in China in 2002 and established on commercial plantations in 2003. Operational application of GMO's in forestry depends on technical, economical, political and public aspects, but the development of adequate regulatory frameworks and public acceptance of transgenic trees will define the future of this technology in forestry.

Sofía, Valenzuela; Claudio, Balocchi; Jaime, Rodríguez.

2006-06-01

284

Expression Systems and Species Used for Transgenic Animal Bioreactors  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Transgenic animal bioreactors can produce therapeutic proteins with high value for pharmaceutical use. In this paper, we compared different systems capable of producing therapeutic proteins (bacteria, mammalian cells, transgenic plants, and transgenic animals) and found that transgenic animals were potentially ideal bioreactors for the synthesis of pharmaceutical protein complexes. Compared with other transgenic animal expression systems (egg white, blood, urine, seminal plasma, and silkworm ...

Yanli Wang; Sihai Zhao; Liang Bai; Jianglin Fan; Enqi Liu

2013-01-01

285

Transgenic Crops: An Introduction and Resource Guide  

Science.gov (United States)

Developed by four professors in the Soil and Crops Sciences and Life Sciences Departments at Colorado State University, this site aims to "provide balanced information and links to other resources on the technology and issues surrounding transgenic crops (also known as genetically modified or GM crops)." None of the authors is affiliated with companies involved in transgenic crop development or with groups campaigning against such crops. The site covers topics such as plant breeding, how transgenic crops are made -- including a Flash demo (not working at time of review), regulation of transgenic crops, current and future transgenic products, risks and concerns, and news updates. The authors deliberately steer clear of the moral or ethical implications of transgenic technology, staying focused on the scientific issues. Throughout the site, links are provided to related sites and other resources. Other sections include a bibliography, quiz, and FAQ.

286

Differential transgene expression in brain cells in vivo and in vitro from AAV-2 vectors with small transcriptional control units  

International Nuclear Information System (INIS)

Adeno-associated- (AAV) based vectors are promising tools for gene therapy applications in several organs, including the brain, but are limited by their small genome size. Two short promoters, the human synapsin 1 gene promoter (hSYN) and the murine cytomegalovirus immediate early promoter (mCMV), were evaluated in bicistronic AAV-2 vectors for their expression profiles in cultured primary brain cells and in the rat brain. Whereas transgene expression from the hSYN promoter was exclusively neuronal, the murine CMV promoter targeted expression mainly to astrocytes in vitro and showed weak transgene expression in vivo in retinal and cortical neurons, but strong expression in thalamic neurons. We propose that neuron specific transgene expression in combination with enhanced transgene capacity will further substantially improve AAV based vector technology

287

Transgenic Spodoptera exigua: possibilities for their use  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Transgenic Spodoptera exigua are being developed by microinjection of a piggyBac vector. The vector expresses green fluorescent protein (GFP) under the control of the actin promoter. Forty percent of the first-instar larvae that hatched from the injected eggs were green fluorescent. However, after backcrossing none of the G1 first-instar larvae was fluorescent and a transgenic line could not be established. Several possibilities for the use of transgenic insects are discussed

Gerritsen, L. J. M.; Visser, J. H.; Jongsma, M. A.

2002-01-01

288

Promoter Sequences for Defining Transgene Expression  

Science.gov (United States)

The design of reverse genetic experiments that utilize transgenic approaches often requires transgenes to be expressed in a predefined pattern and there is limited information regarding the gene expression profile for specific promoters. It is important that expression patterns are predetermined in the specific genotype targeted for transformation because the same promoter-transgene construct can produce different expression patterns in different host species. This chapter compares constitutive, targeted, or inducible promoters that have been characterized in specific cereal species.

Jones, Huw D.; Sparks, Caroline A.

289

Relevance of BAC transgene copy number in mice: transgene copy number variation across multiple transgenic lines and correlations with transgene integrity and expression  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Bacterial artificial chromosomes (BACs) are excellent tools for manipulating large DNA fragments and, as a result, are increasingly utilized to engineer transgenic mice by pronuclear injection. The demand for BAC transgenic mice underscores the need for careful inspection of BAC integrity and fidelity following transgenesis, which may be crucial for interpreting transgene function. Thus, it is imperative that reliable methods for assessing these parameters are available. However, there are li...

Chandler, Kelly J.; Chandler, Ronald L.; Broeckelmann, Eva M.; Hou, Yue; Southard-smith, E. Michelle; Mortlock, Douglas P.

2007-01-01

290

Relative Fitness of Transgenic vs. Non-Transgenic Maize x Teosinte Hybrids: a Field Evalutation  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Concern has been often expressed regarding the impact and persistence of transgenes that enter wild populations via gene flow. The impact of a transgene and its persistence are largely determined by the relative fitness of transgenic hybrids and hybrid derivatives compared to non-transgenic plants. Nevertheless, few studies have addressed this question experimentally in the field. Despite the economic importance of maize, and the fact that it naturally hybridizes with the teosinte taxon Ze...

Clegg, J.; Ellstrand, N. C.; Guadagnuolo, Roberto

2008-01-01

291

Production of recombinant proteins in milk of transgenic and non-transgenic goats  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Among all the transgenic mammalians produced so far, goats have represented an excellent model of transgenesis when considering the factors such as the market demand for protein, volume of milk produced per lactation and reproductive rate. Various recombinant proteins have been obtained from the transgenic and non-transgenic goats, and among these, human antithrombin, produced by the transgenic goats, was the first recombinant protein of animal origin to be released as a drug for the clinical...

Raylene Ramos Moura; Luciana Magalhães Melo; Vicente José de Figueirêdo Freitas

2011-01-01

292

A Built-In Strategy for Containment of Transgenic Plants: Creation of Selectively Terminable Transgenic Rice  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Plant transgenic technology has been widely utilized for engineering crops for trait improvements and for production of high value proteins such as pharmaceuticals. However, the unintended spreading of commercial transgenic crops by pollination and seed dispersal is a major concern for environmental and food safety. Simple and reliable containment strategies for transgenes are highly desirable. Here we report a novel method for creating selectively terminable transgenic rice. In this method, ...

Lin, Chaoyang; Fang, Jun; Xu, Xiaoli; Zhao, Te; Cheng, Jiaan; Tu, Juming; Ye, Gongyin; Shen, Zhicheng

2008-01-01

293

213Bi (?-Emitter)–Antibody Targeting of Breast Cancer Metastases in the neu-N Transgenic Mouse Model  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Treatment failure in breast cancer is largely the failure to control metastatic dissemination. In this study, we investigated the efficacy of an antibody against the rat variant of HER-2/neu, labeled with the ?-particle emitter 213Bi to treat widespread metastases in a rat/neu transgenic mouse model of metastatic mammary carcinoma. The model manifests wide-spread dissemination of tumor cells leading to osteolytic bone lesions and liver metastases, common sites of clinical metastases. The max...

Song, Hong; Shahverdi, Karineh; Huso, David L.; Esaias, Caroline; Fox, James; Liedy, Allison; Zhang, Zhe; Reilly, R. Todd; Apostolidis, Christos; Morgenstern, Alfred; Sgouros, George

2008-01-01

294

Combining M-FISH and Quantum Dot technology for fast chromosomal assignment of transgenic insertions  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Physical mapping of transgenic insertions by Fluorescence in situ Hybridization (FISH is a reliable and cost-effective technique. Chromosomal assignment is commonly achieved either by concurrent G-banding or by a multi-color FISH approach consisting of iteratively co-hybridizing the transgenic sequence of interest with one or more chromosome-specific probes at a time, until the location of the transgenic insertion is identified. Results Here we report a technical development for fast chromosomal assignment of transgenic insertions at the single cell level in mouse and rat models. This comprises a simplified 'single denaturation mixed hybridization' procedure that combines multi-color karyotyping by Multiplex FISH (M-FISH, for simultaneous and unambiguous identification of all chromosomes at once, and the use of a Quantum Dot (QD conjugate for the transgene detection. Conclusions Although the exploitation of the unique optical properties of QD nanocrystals, such as photo-stability and brightness, to improve FISH performance generally has been previously investigated, to our knowledge this is the first report of a purpose-designed molecular cytogenetic protocol in which the combined use of QDs and standard organic fluorophores is specifically tailored to assist gene transfer technology.

Yusuf Mohammed

2011-12-01

295

Somatic hypermutation of an immunoglobulin mu heavy chain transgene  

Digital Repository Infrastructure Vision for European Research (DRIVER)

We have analyzed somatic hypermutation of an immunoglobulin (Ig) heavy chain transgene. Hybridomas expressing the transgene were produced from immunized transgenic mice and transgene copies were sequenced to assay for mutation. In two IgM-producing hybridomas, as well as in several IgG-producing hybridomas, mutations were found in the VDJ region of the transgene. In the IgM-producing hybridomas, both mutated and unmutated transgene copies were present and expressed as mRNA. Several mutated tr...

1993-01-01

296

Production of transgenic livestock: promise fulfilled.  

Science.gov (United States)

The introduction of specific genes into the genome of farm animals and its stable incorporation into the germ line has been a major technological advance in agriculture. Transgenic technology provides a method to rapidly introduce "new" genes into cattle, swine, sheep, and goats without crossbreeding. It is a more extreme methodology, but in essence, not really different from crossbreeding or genetic selection in its result. Methods to produce transgenic animals have been available for more than 20 yr, yet recently lines of transgenic livestock have been developed that have the potential to improve animal agriculture and benefit producers and/or consumers. There are a number of methods that can be used to produce transgenic animals. However, the primary method to date has been the microinjection of genes into the pronuclei of zygotes. This method is one of an array of rapidly developing transgenic methodologies. Another method that has enjoyed recent success is that of nuclear transfer or "cloning." The use of this technique to produce transgenic livestock will profoundly affect the use of transgenic technology in livestock production. Cell-based, nuclear transfer or cloning strategies have several distinct advantages for use in the production of transgenic livestock that cannot be attained using pronuclear injection of DNA. Practical applications of transgenesis in livestock production include enhanced prolificacy and reproductive performance, increased feed utilization and growth rate, improved carcass composition, improved milk production and/or composition, and increased disease resistance. One practical application of transgenics in swine production is to improve milk production and/or composition. To address the problem of low milk production, transgenic swine over-expressing the milk protein bovine alpha-lactalbumin were developed and characterized. The outcomes assessed were milk composition, milk yield, and piglet growth. Our results indicate that transgenic overexpression of milk proteins may provide a means to improve swine lactation performance. PMID:15000404

Wheeler, M B

2003-01-01

297

Plant Transformation: Needs and Futurity of the Transgenes  

Directory of Open Access Journals (Sweden)

Full Text Available To produce transgenic plants which have various applications in agricultural and non-agricultural fields, a marker gene is necessary to recover a viable transgenic plant. To express or transcribe of transgenes, utilization of promoters is also unavoidable. Analysis of transgenes includes copy number, insertion site, integration stability, expression and it`s pattern and variability is immensely important in order to develop a successful transgenic event. This review presents the necessities for better recovery of transgenic plants, transcription or expression of transgenes, as well as methods to analyze transgenes.

Behrooz Darbani

2008-01-01

298

Nontarget DNA sequences reduce the transgene length necessary for RNA-mediated tospovirus resistance in transgenic?plants  

Digital Repository Infrastructure Vision for European Research (DRIVER)

RNA-mediated virus resistance has recently been shown to be the result of post-transcriptional transgene silencing in transgenic plants. This study was undertaken to characterize the effect of transgene length and nontarget DNA sequences on RNA-mediated tospovirus resistance in transgenic plants. Transgenic Nicotiana benthamiana plants were generated to express different regions of the nucleocapsid (N) protein of tomato spotted wilt (TSWV) tospovirus. Transgenic plants expressing half-gene se...

Pang, Sheng-zhi; Jan, Fuh-jyh; Gonsalves, Dennis

1997-01-01

299

BAC Transgenic Mice Reveal Distal Cis-Regulatory Elements Governing BDNF Gene Expression  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Brain-derived neurotrophic factor (BDNF), a member of the neurotrophin family of neurotrophic factors, has important functions in the peripheral and central nervous system of vertebrates. We have generated bacterial artificial chromosome (BAC) transgenic mice harboring 207 kb of the rat BDNF (rBDNF) locus containing the gene, 13 kb of genomic sequences upstream of BDNF exon I, and 144 kb downstream of protein encoding exon IX, in which protein coding region was replaced with the lacZ reporter...

Koppel, Indrek; Aid-pavlidis, Tamara; Jaanson, Kaur; Sepp, Mari; Palm, Kaia; Timmusk, To?nis

2010-01-01

300

The Renin-Angiotensin System Influences Ocular Endothelial Cell Proliferation in Diabetes : Transgenic and Interventional Studies  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Neovascularization in the retina and iris of diabetic patients is a major cause of severe visual loss. However, study of these lesions is compromised by the lack of a comparable diabetic rodent model. Because the vasoactive and angiogenic agent, angiotensin II, is involved in diabetic microvascular disease, we aimed to determine whether endothelial cell proliferation could be induced in the retinae and irides of hypertensive transgenic (mRen-2)27 rats that display an enhanced extra-renal reni...

Moravski, Christina J.; Skinner, Sandford L.; Stubbs, Anthony J.; Sarlos, Stella; Kelly, Darren J.; Cooper, Mark E.; Gilbert, Richard E.; Wilkinson-berka, Jennifer L.

2003-01-01

 
 
 
 
301

Mitochondrial degradation by autophagy (mitophagy) in GFP-LC3 transgenic hepatocytes during nutrient deprivation  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Fasting in vivo and nutrient deprivation in vitro enhance sequestration of mitochondria and other organelles by autophagy for recycling of essential nutrients. Here our goal was to use a transgenic mouse strain expressing green fluorescent protein (GFP) fused to rat microtubule-associated protein-1 light chain 3 (LC3), a marker protein for autophagy, to characterize the dynamics of mitochondrial turnover by autophagy (mitophagy) in hepatocytes during nutrient deprivation. In complete growth m...

Kim, Insil; Lemasters, John J.

2010-01-01

302

A simplified method of generating transgenic Xenopus  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Currently transgenic frog embryos are generated using restriction-enzyme-mediated integration (REMI) on decondensed sperm nuclei followed by nuclear transplantation into unfertilized eggs. We have developed a simplified version of this protocol that has the potential to increase the numbers of normally developing transgenic embryos.

Sparrow, Duncan B.; Latinkic, Branko; Mohun, Tim J.

2000-01-01

303

THE USE OF TRANSGENES FOR WEED MANAGEMENT  

Science.gov (United States)

During the ten years of the availability of commercial, transgenic crops, herbicide resistance has been the most important transgenically conferred crop trait. At this time, almost all of these crops are glyphosate-resistant soybean, maize, cotton, or canola. Bromoxynil-resistant crops are no long...

304

[Progress in transgenic fish techniques and application].  

Science.gov (United States)

Transgenic technique provides a new way for fish breeding. Stable lines of growth hormone gene transfer carps, salmon and tilapia, as well as fluorescence protein gene transfer zebra fish and white cloud mountain minnow have been produced. The fast growth characteristic of GH gene transgenic fish will be of great importance to promote aquaculture production and economic efficiency. This paper summarized the progress in transgenic fish research and ecological assessments. Microinjection is still the most common used method, but often resulted in multi-site and multi-copies integration. Co-injection of transposon or meganuclease will greatly improve the efficiency of gene transfer and integration. "All fish" gene or "auto gene" should be considered to produce transgenic fish in order to eliminate misgiving on food safety and to benefit expression of the transferred gene. Environmental risk is the biggest obstacle for transgenic fish to be commercially applied. Data indicates that transgenic fish have inferior fitness compared with the traditional domestic fish. However, be-cause of the genotype-by-environment effects, it is difficult to extrapolate simple phenotypes to the complex ecological interactions that occur in nature based on the ecological consequences of the transgenic fish determined in the laboratory. It is critical to establish highly naturalized environments for acquiring reliable data that can be used to evaluate the environ-mental risk. Efficacious physical and biological containment strategies remain to be crucial approaches to ensure the safe application of transgenic fish technology. PMID:21586396

Ye, Xing; Tian, Yuan-Yuan; Gao, Feng-Ying

2011-05-01

305

Endogenous immunoglobulin expression in mu transgenic mice.  

Science.gov (United States)

Transgenic mice (M54) containing a functional mu heavy chain were examined to determine the effects of the transgene on rearrangement and expression of endogenous immunoglobulin genes. Two major novel findings are presented. (i) In transgenic mice, the expressed endogenous VH repertoire in LPS-generated B cell blasts and hybridomas is skewed toward expression of JH-proximal VH families (VH7183 and Q52). (ii) There is an increase in the frequency of B cells expressing lambda light chain genes in transgenic mice. Furthermore, in Abelson-MuLV transformed pre-B cells, VH to DJH is inhibited more than the D to JH rearrangement. The results presented indicate that the transgene skews the expressed VH repertoire by inhibiting the VH to DJH rearrangement while permitting an expansion of B cells expressing limited VH and lambda light chain genes. PMID:1902746

Iacomini, J; Yannoutsos, N; Bandyopadhay, S; Imanishi-Kari, T

1991-02-01

306

Segregation of transgenes in maize.  

Science.gov (United States)

Progeny recovered from backcrossed transgenic maize tissue culture regenerants (R0) were analyzed to determine the segregation, expression, and stability of the introduced genes. Transgenic A188 x B73 R0 plants (regenerated from embryogenic suspension culture cells transformed by microprojectile bombardment; see [9]) were pollinated with nontransformed B73 pollen. Inheritance of a selectable marker gene, bar, and a nonselectable marker gene, uidA, was analyzed in progeny (R1) representing four independent transformation events. Activity of the bar gene product, phosphinothricin acetyltransferase (PAT), was assessed in plants comprising the four R1 populations. The number of R1 plants containing PAT activity per total number of R1 plants recovered for each population was 2/7, 19/34, 3/14 and 73/73. Molecular analysis confirmed the segregation of bar in three R1 populations and the lack of segregation in one R1 population. Cosegregation analysis indicated genetic linkage of bar and uidA in all four R1 populations. Analysis of numerous R2 plants derived from crossing transformed R1 plants with nontransformed inbreds revealed 1:1 segregation of PAT activity in three of four lines, including the line that failed to segregate in the R1 generation. Integrated copies of bar in one line appeared to be unstable or poorly transmitted. PMID:1731983

Spencer, T M; O'Brien, J V; Start, W G; Adams, T R; Gordon-Kamm, W J; Lemaux, P G

1992-01-01

307

Glyphostate-drift but not herbivory alters the rate of transgene flow from single and stacked trait transgenic canola (Brassica napus L.) to non-transgenic B. napus and B. rapa  

Science.gov (United States)

While transgenic plants can offer agricultural benefits, the escape of transgenes out of crop fields is a major environmental concern. Escape of transgenic herbicide resistance has occurred between transgenic Brassica napus (canola) and weedy species in numerous locations. In t...

308

Rapid characterization of transgenic and non-transgenic soybean oils by chemometric methods using NIR spectroscopy  

Science.gov (United States)

Near infrared (NIR) spectroscopy and multivariate classification were applied to discriminate soybean oil samples into non-transgenic and transgenic. Principal Component Analysis (PCA) was applied to extract relevant features from the spectral data and to remove the anomalous samples. The best results were obtained when with Support Vectors Machine-Discriminant Analysis (SVM-DA) and Partial Least Squares-Discriminant Analysis (PLS-DA) after mean centering plus multiplicative scatter correction. For SVM-DA the percentage of successful classification was 100% for the training group and 100% and 90% in validation group for non transgenic and transgenic soybean oil samples respectively. For PLS-DA the percentage of successful classification was 95% and 100% in training group for non transgenic and transgenic soybean oil samples respectively and 100% and 80% in validation group for non transgenic and transgenic respectively. The results demonstrate that NIR spectroscopy can provide a rapid, nondestructive and reliable method to distinguish non-transgenic and transgenic soybean oils.

Luna, Aderval S.; da Silva, Arnaldo P.; Pinho, Jéssica S. A.; Ferré, Joan; Boqué, Ricard

309

Tissue-specific and hormonal regulation of calbindin-D9K fusion genes in transgenic mice.  

Science.gov (United States)

The rat Calbindin-D9K (CaBP9K) gene is mainly expressed in intestine, uterus, and lung and is regulated in a complex tissue-specific manner. To analyze the role of potential regulatory elements, previously defined by DNaseI hypersensivity, we made transgenic mice containing truncated rat CaBP9K fusion gene with simian virus 40 large T antigen and the chloramphenicol acetyltransferase as reporter genes. The transgenes contained CaBP9K promoter fragments with 5' end points at -4400, -1011, and -117 base pairs (bp), whereas the 3' end points was at +365 bp. Northern blot analysis of T antigen expression and chloramphenicol acetyltransferase enzyme-linked immunosorbent assay indicated that a positive element, probably the distal intestine-specific DNaseI HS, necessary to target the expression of the transgene in the intestine, is present between -4400 and -1011 bp. The cephalo-caudal gradient of expression of the transgene along the small intestine was similar to those of the endogenous gene, but an ectopic expression of the transgene was observed in the colon. The -1011 transgene was expressed in epithelial alveolar cells of the lung, in renal proximal tubule cells, and in uterine myometrium, as judged from immunocytochemical, histological, and Northern blot analyses. The shortest, -117 construct was only expressed in uterine myometrium, and it was under a strict estrogen dependence like the endogenous gene. Finally, responsiveness to vitamin D in the duodenum was observed with the largest, -4400 construct. Thus, different tissues utilize distinct cis-acting elements to direct and regulate the expression of the rat CaBP9K gene. PMID:8663193

Romagnolo, B; Cluzeaud, F; Lambert, M; Colnot, S; Porteu, A; Molina, T; Tomasset, M; Vandewalle, A; Kahn, A; Perret, C

1996-07-12

310

Disease-associated polymorphisms in ERAP1 do not alter endoplasmic reticulum stress in patients with ankylosing spondylitis.  

Science.gov (United States)

The mechanism by which human leukocyte antigen B27 (HLA-B27) contributes to ankylosing spondylitis (AS) remains unclear. Genetic studies demonstrate that association with and interaction between polymorphisms of endoplasmic reticulum aminopeptidase 1 (ERAP1) and HLA-B27 influence the risk of AS. It has been hypothesised that ERAP1-mediated HLA-B27 misfolding increases endoplasmic reticulum (ER) stress, driving an interleukin (IL) 23-dependent, pro-inflammatory immune response. We tested the hypothesis that AS-risk ERAP1 variants increase ER-stress and concomitant pro-inflammatory cytokine production in HLA-B27(+) but not HLA-B27(-) AS patients or controls. Forty-nine AS cases and 22 healthy controls were grouped according to HLA-B27 status and AS-associated ERAP1 rs30187 genotypes: HLA-B27(+)ERAP1(risk), HLA-B27(+)ERAP1(protective), HLA-B27(-)ERAP1(risk) and HLA-B27(-)ERAP1(protective). Expression levels of ER-stress markers GRP78 (8?kDa glucose-regulated protein), CHOP (C/EBP-homologous protein) and inflammatory cytokines were determined in peripheral blood mononuclear cell and ileal biopsies. We found no differences in ER-stress gene expression between HLA-B27(+) and HLA-B27(-) cases or healthy controls, or between cases or controls stratified by carriage of ERAP1 risk or protective alleles in the presence or absence of HLA-B27. No differences were observed between expression of IL17A or TNF (tumour necrosis factor) in HLA-B27(+)ERAP1(risk), HLA-B27(+)ERAP1(protective) and HLA-B27(-)ERAP1(protective) cases. These data demonstrate that aberrant ERAP1 activity and HLA-B27 carriage does not alter ER-stress levels in AS, suggesting that ERAP1 and HLA-B27 may influence disease susceptibility through other mechanisms. PMID:25354578

Kenna, T J; Lau, M C; Keith, P; Ciccia, F; Costello, M-E; Bradbury, L; Low, P-L; Agrawal, N; Triolo, G; Alessandro, R; Robinson, P C; Thomas, G P; Brown, M A

2015-01-01

311

Generation of transgenic Hydra by embryo microinjection.  

Science.gov (United States)

As a member of the phylum Cnidaria, the sister group to all bilaterians, Hydra can shed light on fundamental biological processes shared among multicellular animals. Hydra is used as a model for the study of regeneration, pattern formation, and stem cells. However, research efforts have been hampered by lack of a reliable method for gene perturbations to study molecular function. The development of transgenic methods has revitalized the study of Hydra biology(1). Transgenic Hydra allow for the tracking of live cells, sorting to yield pure cell populations for biochemical analysis, manipulation of gene function by knockdown and over-expression, and analysis of promoter function. Plasmid DNA injected into early stage embryos randomly integrates into the genome early in development. This results in hatchlings that express transgenes in patches of tissue in one or more of the three lineages (ectodermal epithelial, endodermal epithelial, or interstitial). The success rate of obtaining a hatchling with transgenic tissue is between 10% and 20%. Asexual propagation of the transgenic hatchling is used to establish a uniformly transgenic line in a particular lineage. Generating transgenic Hydra is surprisingly simple and robust, and here we describe a protocol that can be easily implemented at low cost. PMID:25285460

Juliano, Celina E; Lin, Haifan; Steele, Robert E

2014-01-01

312

G2R Cre Reporter Transgenic Zebrafish  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The Cre/loxP site-specific recombination system has been widely used to manipulate DNA in vivo and to study gene function in the mouse by inducible transgenic and conditional gene targeting. To fully use this powerful genetic tool in a relatively new animal model, zebrafish, we generated reporter transgenic lines for easy detection of Cre recombinase activity in vivo. The transgenic fish lines, designated G2R, express two fluorescent protein genes, GFP and RFP, under the control of the ubiqui...

Yoshikawa, Shunichi; Kawakami, Koichi; Zhao, Xinping C.

2008-01-01

313

Expression of multiple proteins in transgenic plants  

Science.gov (United States)

A method is disclosed for the production of multiple proteins in transgenic plants. A DNA construct for introduction into plants includes a provision to express a fusion protein of two proteins of interest joined by a linking domain including plant ubiquitin. When the fusion protein is produced in the cells of a transgenic plant transformed with the DNA construction, native enzymes present in plant cells cleave the fusion protein to release both proteins of interest into the cells of the transgenic plant. Since the proteins are produced from the same fusion protein, the initial quantities of the proteins in the cells of the plant are approximately equal.

Vierstra, Richard D. (Madison, WI); Walker, Joseph M. (Madison, WI)

2002-01-01

314

LKB1 deletion with the RIP2.Cre transgene modifies pancreatic ?-cell morphology and enhances insulin secretion in vivo  

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The tumor suppressor liver kinase B1 (LKB1), also called STK11, is a protein kinase mutated in Peutz-Jeghers syndrome. LKB1 phosphorylates AMP-activated protein kinase (AMPK) and several related protein kinases. Whereas deletion of both catalytic isoforms of AMPK from the pancreatic ?-cell and hypothalamic neurons using the rat insulin promoter (RIP2).Cre transgene (?AMPKdKO) diminishes insulin secretion in vivo, deletion of LKB1 in the ?-cell with an inducible Pdx-1.CreER transgene enhanc...

Sun, Gao; Tarasov, Andrei I.; Mcginty, James A.; French, Paul M.; Mcdonald, Angela; Leclerc, Isabelle; Rutter, Guy A.

2010-01-01

315

Transgene-specific host responses in cutaneous gene therapy: the role of cells expressing the transgene  

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A major issue in long-term gene therapy is host immune responses to therapeutic cells when transgene encodes a potential antigen. The nature of these responses depends on several factors including the type of cell and tissue expressing the transgene. Keratinocytes and fibroblasts, which are known to display distinct immunogenic profiles, are both potential targets for transgene expression in cutaneous gene therapy. However, whether there is an immunological advantage in targeting one cell typ...

Zhang, Zhenghua; Kuscu, Cem; Ghazizadeh, Soosan

2009-01-01

316

Comparison of nutritional value of transgenic peanut expressing bar and rcg3 genes with non-transgenic counterparts  

International Nuclear Information System (INIS)

The transgenic peanut plants expressing bar and rcg3 genes were subjected to assessment of any change in nutritional value of the crop at various locations. The protein and fat contents of transgenic lines were compared with the non-transgenic parent varieties. Protein content in the transgenic lines was higher as compared to that in non-transgenic counterparts and differences among locations for fat and protein content were significant. No differences among fatty acids were recorded for genes, events and locations. Irrespective of small differences, all the values were in range described for this crop and transgenic lines appeared to be substantially equivalent to non-transgenic parent varieties. (author)

317

Mutagenicity of comfrey (Symphytum Officinale) in rat liver  

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Comfrey is a rat liver toxin and carcinogen that has been used as a vegetable and herbal remedy by humans. In order to evaluate the mechanisms underlying its carcinogenicity, we examined the mutagenicity of comfrey in the transgenic Big Blue rat model. Our results indicate that comfrey is mutagenic in rat liver and the types of mutations induced by comfrey suggest that its tumorigenicity results from the genotoxicity of pyrrolizidine alkaloids in the plant.

Mei, N.; Guo, L.; Fu, P. P.; Heflich, R. H.; Chen, T.

2005-01-01

318

Mutagenicity of comfrey (Symphytum Officinale) in rat liver.  

Science.gov (United States)

Comfrey is a rat liver toxin and carcinogen that has been used as a vegetable and herbal remedy by humans. In order to evaluate the mechanisms underlying its carcinogenicity, we examined the mutagenicity of comfrey in the transgenic Big Blue rat model. Our results indicate that comfrey is mutagenic in rat liver and the types of mutations induced by comfrey suggest that its tumorigenicity results from the genotoxicity of pyrrolizidine alkaloids in the plant. PMID:15726100

Mei, N; Guo, L; Fu, P P; Heflich, R H; Chen, T

2005-03-14

319

Accumulation of nickel in transgenic tobacco  

Science.gov (United States)

The accumulation of heavy metal Ni in the roots and leaves of four T1 transgenic lines of tobacco (T(1)20E, T(1)24C, T(1)18B1 and T(1)20B) expressing eiMT1 from E.indica was assessed. The aim of the study was to investigate the level of Ni accumulation in the leaves and roots of each transgenic lines and to evaluate the eligibility of the plants to be classified as a phytoremediation agent. All of the transgenic lines showed different ability in accumulating different metals and has translocation factor (TF) less than 1 (TFtransgenic lines, transgenic line T(1)24C showed the highest accumulation of Ni (251.9 ± 0.014 mg/kg) and the lowest TF value (TFT(1)24C=0.0875) at 60 ppm Ni.

Sidik, Nik Marzuki; Othman, Noor Farhan

2013-11-01

320

AN APPROACH TO TRANSGENIC CROP MONITORING  

Science.gov (United States)

Remote sensing by aerial or satellite images may provide a method of identifying transgenic pesticidal crop distribution in the landscape. Genetically engineered crops containing bacterial gene(s) that express an insecticidal protein from Bacillus thuringiensis (Bt) are regulated...

 
 
 
 
321

Phytochrome transgenics: functional, ecological and biotechnological applications.  

Science.gov (United States)

The phytochromes have important functions in regulating plant growth and development in response to signals perceived from the natural light environment. In particular, the phytochrome-mediated shade avoidance syndrome has major significance for competition between plants growing in natural dense communities. In recent years, the availability of DNA sequences coding for members of the phytochrome family has enabled the construction of transgenic plants that express these sequences to high levels. Introduced PHY genes expressed in heterologous or homologous hosts yield apoproteins that combine with chromophores and are physiologically functional. Physiological analysis of transgenic plants expressing introduced PHYA and PHYB coding sequences has contributed to understanding the functions of phytochromes A and B. Ecological experiments with transgenic PHYA expressers have provided a novel test of the adaptive plasticity hypothesis, and point the way to a transgenic programme to improve crop plants. PMID:7881071

Smith, H

1994-10-01

322

Transgenic Wheat, Barley and Oats: Future Prospects  

Science.gov (United States)

Following the success of transgenic maize and rice, methods have now been developed for the efficient introduction of genes into wheat, barley and oats. This review summarizes the present position in relation to these three species, and also uses information from field trial databases and the patent literature to assess the future trends in the exploitation of transgenic material. This analysis includes agronomic traits and also discusses opportunities in expanding areas such as biofuels and biopharming.

Dunwell, Jim M.

323

Transgenic animals and their application in medicine  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Transgenic animals are animals that are genetically altered to have traits that mimic symptoms of specific human pathologies. They provide genetic models of various human diseases which are important in understanding disease and developing new targets. In early 1980 Gordon and co-workers described the first gene addition experiment using the microinjection technology and since then the impact of transgenic technology on basic research has been significant. Within 20 years of its inception, AT...

Bagle Tr, Kunkulol Rr

2013-01-01

324

Brain changes in endothelin-1 transgenic mice  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The purpose of this study was to investigate the effect of endothelin-1 overexpression in the central nervous system of endothelin-1 transgenic NMRI mice. The brains were embedded in paraffin and examined with immunohistochemical methods. Using LacZ as a marker it was possible to show endothelin transgene expression in neurons of the brainstem, the piriform cortex, the nuclei of the hypothalamus and thalamus, the frontal cortex, the nuclei of the cerebellum, to a lesser extent also in neurons...

Scha?fer, Silke

2010-01-01

325

A Transgenic Tri-Modality Reporter Mouse  

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Transgenic mouse with a stably integrated reporter gene(s) can be a valuable resource for obtaining uniformly labeled stem cells, tissues, and organs for various applications. We have generated a transgenic mouse model that ubiquitously expresses a tri-fusion reporter gene (fluc2-tdTomato-ttk) driven by a constitutive chicken ?-actin promoter. This “Tri-Modality Reporter Mouse” system allows one to isolate most cells from this donor mouse and image them for bioluminescent (fluc2), fluore...

Yan, Xinrui; Ray, Pritha; Paulmurugan, Ramasamy; Tong, Ricky; Gong, Yongquan; Sathirachinda, Ataya; Wu, Joseph C.; Gambhir, Sanjiv S.

2013-01-01

326

Transgene flow: Facts, speculations and possible countermeasures.  

Science.gov (United States)

Convincing evidence has accumulated that unintended transgene escape occurs in oilseed rape, maize, cotton and creeping bentgrass. The escaped transgenes are found in variant cultivars, in wild type plants as well as in hybrids of sexually compatible species. The fact that in some cases stacked events are present that have not been planted commercially, implies unintended recombination of transgenic traits. As the consequences of this continuous transgene escape for the ecosystem cannot be reliably predicted, I propose to use more sophisticated approaches of gene technology in future. If possible GM plants should be constructed using either site-directed mutagenesis or cisgenic strategies to avoid the problem of transgene escape. In cases where a transgenic trait is needed, efficient containment should be the standard approach. Various strategies available or in development are discussed. Such a cautious approach in developing novel types of GM crops will enhance the sustainable potential of GM crops and thus increase the public trust in green gene technology. PMID:25523171

Ryffel, Gerhart U

2014-10-01

327

TRANSGENIC FISH MODEL IN ENVIRONMENTAL TOXICOLOGY  

Directory of Open Access Journals (Sweden)

Full Text Available A number of experiments and the use of drugs have been performed in fish. The fish may be used as model organism in various biological experiments, including environmental toxicology. Aquatic animals are being engineered to increase aquaculture production, for medical and industrial research, and for ornamental reasons. Fish have been found to play an important role in assessing potential risks associated with exposure to toxic substances in aquatic environment. Hence, it has been thought that the development of transgenic fish can enhance the use of fish in environmental toxicology. India has developed experimental transgenics of rohu fish, zebra fish, cat fish and singhi fish. Genes, promoters and vectors of indigenous origin are now available for only two species namely rohu and singhi for engineering growth. Development of fish model carrying identical transgenes to those found in rodents is beneficial and has shown that several aspects of in vivo mutagenesis are similar between the two classes of vertebrates. Fish shows the frequencies of spontaneous mutations similar to rodents and respond to mutagen exposure consistent with known mutagenic mechanisms. The feasibility of in vivo mutation analysis using transgenic fish has been demonstrated and the potential value of transgenic fish as a comparative animal model has been illustrated. Therefore, the transgenic fish can give the significant contribution to study the environmental toxicity in animals as a whole.

Madhuri Sharma

2012-05-01

328

Sustained, localized transgene expression mediated from lentivirus-loaded biodegradable polyester elastomers.  

Science.gov (United States)

The study of biomaterials for gene delivery in tissue engineering and regenerative medicine is a growing area, necessitating the investigation of new biomaterials and gene delivery vectors. Poly(1,8-octanediol citrate) (POC) and poly(glycerol-sebacate) (PGS) are biodegradable, biocompatible elastomers that have tunable mechanical properties, surface characteristics, and degradation rate. The objective of this work was to investigate whether POC and PGS would support the immobilization and release of lentivirus to allow sustained and localized transgene expression. Porous biomaterials were prepared using salt as a porogen, and in vitro and in vivo transgene expression from immobilized and released lentiviruses were assessed. Cells seeded onto biomaterials loaded with lentiviruses yielded titer-dependent transgene expression in vitro. Lentivirus activity on both biomaterials was maintained for at least 5 days. When implanted subcutaneously in rats, POC and PGS with immobilized lentivirus exhibited sustained and localized transgene expression for at least 5 weeks. This research demonstrates that lentivirus immobilization on POC and PGS is feasible and potentially useful for a variety of tissue engineering and regenerative medicine applications. PMID:23065823

Jen, Michele C; Baler, Kevin; Hood, Ashleigh R; Shin, Seungjin; Shea, Lonnie D; Ameer, Guillermo A

2013-05-01

329

The rat as an animal model of Alzheimer's disease  

DEFF Research Database (Denmark)

As a disease model, the laboratory rat has contributed enormously to neuroscience research over the years. It has also been a popular animal model for Alzheimer's disease but its popularity has diminished during the last decade, as techniques for genetic manipulation in rats have lagged behind that of mice. In recent years, the rat has been making a comeback as an Alzheimer's disease model and the appearance of increasing numbers of transgenic rats will be a welcome and valuable complement to the existing mouse models. This review summarizes the contributions and current status of the rat as an animal model of Alzheimer's disease.

Benedikz, Eirikur; Kloskowska, Ewa

2009-01-01

330

Transgenic technologies to induce sterility  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract The last few years have witnessed a considerable expansion in the number of tools available to perform molecular and genetic studies on the genome of Anopheles mosquitoes, the vectors of human malaria. As a consequence, knowledge of aspects of the biology of mosquitoes, such as immunity, reproduction and behaviour, that are relevant to their ability to transmit disease is rapidly increasing, and could be translated into concrete benefits for malaria control strategies. Amongst the most important scientific advances, the development of transgenic technologies for Anopheles mosquitoes provides a crucial opportunity to improve current vector control measures or design novel ones. In particular, the use of genetic modification of the mosquito genome could provide for a more effective deployment of the sterile insect technique (SIT against vector populations in the field. Currently, SIT relies on the release of radiation sterilized males, which compete with wild males for mating with wild females. The induction of sterility in males through the genetic manipulation of the mosquito genome, already achieved in a number of other insect species, could eliminate the need for radiation and increase the efficiency of SIT-based strategies. This paper provides an overview of the mechanisms already in use for inducing sterility by transgenesis in Drosophila and other insects, and speculates on possible ways to apply similar approaches to Anopheles mosquitoes.

Wimmer Ernst A

2009-11-01

331

Generation of stable Xenopus laevis transgenic lines expressing a transgene controlled by weak promoters.  

Science.gov (United States)

Combining two existing protocols of trangenesis, namely the REMI and the I-SceI meganuclease methods, we generated Xenopus leavis expressing a transgene under the control of a promoter that presented a restricted pattern of activity and a low level of expression. This was realized by co-incubating sperm nuclei, the I-SceI enzyme and the transgene prior to transplantation into unfertilized eggs. The addition of the woodchuck hepatitis virus posttranscriptional regulatory element in our constructs further enhanced the expression of the transgene without affecting the tissue-specificity of the promoter activity. Using this combination of methods we produced high rates of fully transgenic animals that stably transmitted the transgene to the next generations with a transmission rate of 50% indicating a single integration event. PMID:19404763

L'hostis-Guidet, Anne; Recher, Gaëlle; Guillet, Brigitte; Al-Mohammad, Abdulrahim; Coumailleau, Pascal; Tiaho, François; Boujard, Daniel; Madigou, Thierry

2009-10-01

332

Rat aquaporin-5 4.3-kb 5?-flanking region differentially regulates expression in salivary gland and lung in vivo  

Digital Repository Infrastructure Vision for European Research (DRIVER)

We previously cloned a 4.3-kb genomic fragment encompassing 5?-flanking regulatory elements of rat aquaporin-5 (Aqp5) that demonstrated preferential transcriptional activity in lung and salivary cells in vitro. To investigate the ability of Aqp5 regulatory elements to direct transgene expression in vivo, transgenic (TG) mice and rats were generated in which the 4.3-kb Aqp5 fragment directed the expression of enhanced green fluorescent protein (EGFP). RT-PCR revealed relative promoter specif...

Zhou, Beiyun; Ann, David K.; Flodby, Per; Minoo, Parviz; Liebler, Janice M.; Crandall, Edward D.; Borok, Zea

2008-01-01

333

Lectin cDNA and transgenic plants derived therefrom  

Science.gov (United States)

Transgenic plants containing cDNA encoding Gramineae lectin are described. The plants preferably contain cDNA coding for barley lectin and store the lectin in the leaves. The transgenic plants, particularly the leaves exhibit insecticidal and fungicidal properties.

Raikhel, Natasha V. (Okemos, MI)

2000-10-03

334

Increased cytokine-induced cytotoxicity of pancreatic islet cells from transgenic mice expressing the Src-like tyrosine kinase GTK.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

BACKGROUND: The loss of beta cells in type 1 diabetes may involve protein kinases because they control cell growth, differentiation, and survival. Previous studies have revealed that GTK, a Src-like protein tyrosine kinase expressed in beta cells (also named Bsk/Iyk), regulates multiple responses including growth and survival of rat insulinoma cells (RINm5F) and differentiation of neuronal PC12 cells. In the present study, we have generated a transgenic mouse expressing a kinase active GTK mu...

Annere?n, C.; Welsh, M.

2001-01-01

335

Bone Loss in Spondyloarthritis Linked to Protein's Misfolding  

Science.gov (United States)

... Arthritis & Rheumatism, focuses on an immune protein called HLA-B27, a member of a class of molecules that ... known for many years that people who carry HLA-B27 are more susceptible to spondyloarthritis, but they haven’ ...

336

Heredity and Arthritis  

Science.gov (United States)

... between ankylosing spondylitis (AS) and a gene called HLA-B27. Ankylosing spondylitis is an inflammatory arthritis affecting primarily ... than 90% of Caucasian AS patients have the HLA-B27 gene, compared to the approximately 7% of the ...

337

Genetics Home Reference: Ankylosing spondylitis  

Science.gov (United States)

... A variation of the HLA-B gene called HLA-B27 increases the risk of developing ankylosing spondylitis. Although many people with ankylosing spondylitis have the HLA-B27 variation, most people with this version of the ...

338

Ankylosing Spondylitis  

Science.gov (United States)

... associated with susceptibility to ankylosing spondylitis is called HLA-B27. But while most people with ankylosing spondylitis have ... ankylosing spondylitis is one to check for the HLA-B27 gene, which is present in the majority of ...

339

Transgenic cultures: from the economic viewpoint  

Directory of Open Access Journals (Sweden)

Full Text Available The introduction of transgenic seeds for agricultural purposes poses modification to their production, due to the potential for reaching desired characteristics such as greater yield, this being fundamental in an economic environment characterised by open market conditions. However, acceptance of products resulting from genetic engineering is far from becoming a simple process; discussion relating to the predominance of private sector interests, the monopoly of knowledge and the safety of such seeds/food is currently in the spotlight. This article presents the main points of debate regarding adoption of transgenic cultures, contributing to discussion about this topic for Colombia.

Mauricio Mosquera

2011-12-01

340

Generation of BAC transgenic epithelial organoids.  

Science.gov (United States)

Under previously developed culture conditions, mouse and human intestinal epithelia can be cultured and expanded over long periods. These so-called organoids recapitulate the three-dimensional architecture of the gut epithelium, and consist of all major intestinal cell types. One key advantage of these ex vivo cultures is their accessibility to live imaging. So far the establishment of transgenic fluorescent reporter organoids has required the generation of transgenic mice, a laborious and time-consuming process, which cannot be extended to human cultures. Here we present a transfection protocol that enables the generation of recombinant mouse and human reporter organoids using BAC (bacterial artificial chromosome) technology. PMID:24204693

Schwank, Gerald; Andersson-Rolf, Amanda; Koo, Bon-Kyoung; Sasaki, Nobuo; Clevers, Hans

2013-01-01

 
 
 
 
341

Advancing environmental risk assessment for transgenic biofeedstock crops  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Abstract Transgenic modification of plants is a key enabling technology for developing sustainable biofeedstocks for biofuels production. Regulatory decisions and the wider acceptance and development of transgenic biofeedstock crops are considered from the context of science-based risk assessment. The risk assessment paradigm for transgenic biofeedstock crops is fundamentally no different from that of current generation transgenic crops, except that the focus of the assessment must ...

Wolt Jeffrey D

2009-01-01

342

IDENTIFICATION OF ESCAPED TRANSGENIC CREEPING BENTGRASS IN OREGON  

Science.gov (United States)

When transgenic plants are cultivated near wild species that are sexually compatible with the crop, gene flow between the crop and wild plants is possible. A resultant concern is that transgene flow and transgene introgression within wild populations could have unintended ecologi...

343

Maize transgenes containing zein promoters are regulated by opaque2  

Science.gov (United States)

Transgenes have great potential in crop improvement, but relatively little is known about the epistatic interaction of transgenes with the native genes in the genome. Understanding these interactions is critical for predicting the response of transgenes to different genetic backgrounds and environm...

344

The substantive equivalence of transgenic (Bt and Chi) and non-transgenic cotton based on metabolite profiles.  

Science.gov (United States)

Compositional studies comparing transgenic with non-transgenic counterpart plants are almost universally required by governmental regulatory bodies. In the present study, two T(2) transgenic cotton lines containing chitinase (Line 11/57) and Bt lines (Line 61) were compared with non-transgenic counterpart. To do this, biochemical characteristics of leaves and seeds, including amino acids, fatty acids, carbohydrates, anions, and cations contents of the studied lines were analyzed using GC/MS, high-performance liquid chromatography (HPLC), and ion chromatography (IC) analyzers, respectively. polymerase chain reaction (PCR) and Western blot analyses confirmed the presence and expression of Chi and Bt genes in the studied transgenic lines. Although, compositional analysis of leaves contents confirmed no significant differences between transgenic and non-transgenic counterpart lines, but it was shown that glucose content of chitinase lines, fructose content of transgenic lines (Bt and chitinase) and asparagine and glutamine of chitinase lines were significantly higher than the non-transgenic counterpart plants. Both the transgenic lines (Bt and chitinase) showed significant decrease in the amounts of sodium in comparison to the non-transgenic counterpart plants. The experiments on the seeds showed that histidine, isoleucine, leucine, and phenylalanine contents of all transgenic and non-transgenic lines were the same, whereas other amino acids were significantly increased in the transgenic lines. Surprisingly, it was observed that the concentrations of stearic acid, myristic acid, oleic acid, and linoleic acid in the chitinase line were significantly different than those of non-transgenic counterpart plants, but these components were the same in both Bt line and its non-transgenic counterpart. It seems that more changes observed in the seed contents than leaves is via this point that seeds are known as metabolites storage organs, so they show greater changes in the metabolites contents comparing to the leaves. PMID:24374853

Modirroosta, Bentol Hoda; Tohidfar, Masoud; Saba, Jalal; Moradi, Foad

2014-03-01

345

In situ methods to localize transgenes and transcripts in interphase nuclei: a tool for transgenic plant research  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Genetic engineering of commercially important crops has become routine in many laboratories. However, the inability to predict where a transgene will integrate and to efficiently select plants with stable levels of transgenic expression remains a limitation of this technology. Fluorescence in situ hybridization (FISH) is a powerful technique that can be used to visualize transgene integration sites and provide a better understanding of transgene behavior. Studies using FISH to characterize tr...

Santos, Ana Paula; Wegel, Eva; Allen, George C.; Thompson, William F.; Stoger, Eva; Shaw, Peter; Abranches, Rita

2006-01-01

346

Comparative analysis of nutritional compositions of transgenic high iron rice with its non-transgenic counterpart.  

Science.gov (United States)

Iron is an essential micronutrient for human nutrition and polished rice contains very low amount of iron. Rice with high iron content in seed endosperm has been developed by insertion of soybean ferritin gene under the control of the endosperm specific glutelin promoter into the genome of indica rice line IR68144. The nutritional composition of the brown and milled rice grain has been compared with that of the non-transgenic rice of the same variety. In this study, the nutritional components, as well as the anti-nutrient levels, were measured. Our studies established that apart from the increased level of iron and zinc in transgenic seeds, the nutritional quality of both the brown and milled rice grains from the transgenic line was substantially equivalent to that of the non-transgenic rice plants. The result clearly shows that the measured amounts of the nutritional components are well within the range of values reported for other commercial lines. PMID:23411185

Gayen, Dipak; Sarkar, Sailendra Nath; Datta, Swapan K; Datta, Karabi

2013-06-01

347

Production of recombinant proteins in milk of transgenic and non-transgenic goats  

Directory of Open Access Journals (Sweden)

Full Text Available Among all the transgenic mammalians produced so far, goats have represented an excellent model of transgenesis when considering the factors such as the market demand for protein, volume of milk produced per lactation and reproductive rate. Various recombinant proteins have been obtained from the transgenic and non-transgenic goats, and among these, human antithrombin, produced by the transgenic goats, was the first recombinant protein of animal origin to be released as a drug for the clinical use in humans. This review reports the aspects inherent to the production of recombinant proteins in the goats, from the production of the animal bioreactors up to the expression of these proteins in their milk.

Raylene Ramos Moura

2011-10-01

348

Mammalian cDNA and prokaryotic reporter sequences silence adjacent transgenes in transgenic mice.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The ovine beta-lactoglobulin gene is expressed efficiently and at high levels in the mammary gland of transgenic mice. In contrast, when this gene is linked to a second gene construct comprising a mammalian cDNA or a CAT reporter sequence it fails to be expressed in the majority of transgenic lines generated. We suggest that mammalian cDNAs and prokaryotic reporter sequences can serve as active foci for gene silencing in the mammalian genome.

Clark, A. J.; Harold, G.; Yull, F. E.

1997-01-01

349

Generation of stable Xenopus laevis transgenic lines expressing a transgene controlled by weak promoters.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Combining two existing protocols of trangenesis, namely the REMI and the I-SceI meganuclease methods, we generated Xenopus leavis expressing a transgene under the control of a promoter that presented a restricted pattern of activity and a low level of expression. This was realized by co-incubating sperm nuclei, the I-SceI enzyme and the transgene prior to transplantation into unfertilized eggs. The addition of the woodchuck hepatitis virus posttranscriptional regulatory element in our constru...

L Hostis-guidet, Anne; Recher, Gae?lle; Guillet, Brigitte; Al-mohammad, Abdulrahim; Coumailleau, Pascal; Tiaho, Franc?ois; Boujard, Daniel; Madigou, Thierry

2009-01-01

350

TRANSGENIC MOUSE MODELS AND PARTICULATE MATTER (PM)  

Science.gov (United States)

The hypothesis to be tested is that metal catalyzed oxidative stress can contribute to the biological effects of particulate matter. We acquired several transgenic mouse strains to test this hypothesis. Breeding of the mice was accomplished by Duke University. Particles employed ...

351

Transgenic cultures: from the economic viewpoint  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The introduction of transgenic seeds for agricultural purposes poses modification to their production, due to the potential for reaching desired characteristics such as greater yield, this being fundamental in an economic environment characterised by open market conditions. However, acceptance of products resulting from genetic en...

Mauricio Mosquera

2011-01-01

352

Transgenic nonhuman primates for neurodegenerative diseases  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Abstract Animal models that represent human diseases constitute an important tool in understanding the pathogenesis of the diseases, and in developing effective therapies. Neurodegenerative diseases are complex disorders involving neuropathologic and psychiatric alterations. Although transgenic and knock-in mouse models of Alzheimer's disease, (AD), Parkinson's disease (PD) and Huntington's disease (HD) have been created, limited representation in clinical aspects has been recognize...

Ws, Chan Anthony

2004-01-01

353

Progress in Xenotransplantation Research Employing Transgenic Pigs  

Directory of Open Access Journals (Sweden)

Full Text Available Microinjection of foreign DNA into pronuclei of a fertilized oocyte has predominantly been used for the generation of transgenic livestock. This technology works reliably, but is inefficient and results in random integration and variable expression patterns in the transgenic offspring. Nevertheless, remarkable achievements have been made with this technology with regard to xenotransplantation. Transgenic pigs that express human complement regulating proteins have been tested in their ability to serve as donors in human organ transplantation (i.e. xenotransplantation. In vitro and in vivo data convincingly show that the hyperacute rejection response can be overcome in a clinically acceptable manner by successfully employing this strategy. The recent developments in nuclear transfer and its merger with the growing genomic data allow targeted and regulatable transgenesis. Systems for efficient homologous recombination in somatic cells are being developed and the first knock-out pigs, carrying a deletion in the a-galactosyltransferase gene, were recently generated. It is anticipated that poly-transgenic pigs will be available as donors for functional xenografts within a few years. Similarly, pigs may serve as donors for a variety of xenogenic cells and tissues. The availability of these technologies is essential to maintain "genetic security" and to ensure absence of unwanted side effects.

H. Niemann

2003-06-01

354

Transgenic Mouse Model of Chronic Beryllium Disease  

Energy Technology Data Exchange (ETDEWEB)

Animal models provide powerful tools for dissecting dose-response relationships and pathogenic mechanisms and for testing new treatment paradigms. Mechanistic research on beryllium exposure-disease relationships is severely limited by a general inability to develop a sufficient chronic beryllium disease animal model. Discovery of the Human Leukocyte Antigen (HLA) - DPB1Glu69 genetic susceptibility component of chronic beryllium disease permitted the addition of this human beryllium antigen presentation molecule to an animal genome which may permit development of a better animal model for chronic beryllium disease. Using FVB/N inbred mice, Drs. Rubin and Zhu, successfully produced three strains of HLA-DPB1 Glu 69 transgenic mice. Each mouse strain contains a haplotype of the HLA-DPB1 Glu 69 gene that confers a different magnitude of odds ratio (OR) of risk for chronic beryllium disease: HLA-DPB1*0401 (OR = 0.2), HLA-DPB1*0201 (OR = 15), HLA-DPB1*1701 (OR = 240). In addition, Drs. Rubin and Zhu developed transgenic mice with the human CD4 gene to permit better transmission of signals between T cells and antigen presenting cells. This project has maintained the colonies of these transgenic mice and tested the functionality of the human transgenes.

Gordon, Terry

2009-05-26

355

Transgenic plants with increased calcium stores  

Science.gov (United States)

The present invention provides transgenic plants over-expressing a transgene encoding a calcium-binding protein or peptide (CaBP). Preferably, the CaBP is a calcium storage protein and over-expression thereof does not have undue adverse effects on calcium homeostasis or biochemical pathways that are regulated by calcium. In preferred embodiments, the CaBP is calreticulin (CRT) or calsequestrin. In more preferred embodiments, the CaBP is the C-domain of CRT, a fragment of the C-domain, or multimers of the foregoing. In other preferred embodiments, the CaBP is localized to the endoplasmic reticulum by operatively associating the transgene encoding the CaBP with an endoplasmic reticulum localization peptide. Alternatively, the CaBP is targeted to any other sub-cellular compartment that permits the calcium to be stored in a form that is biologically available to the plant. Also provided are methods of producing plants with desirable phenotypic traits by transformation of the plant with a transgene encoding a CaBP. Such phenotypic traits include increased calcium storage, enhanced resistance to calcium-limiting conditions, enhanced growth and viability, increased disease and stress resistance, enhanced flower and fruit production, reduced senescence, and a decreased need for fertilizer production. Further provided are plants with enhanced nutritional value as human food or animal feed.

Wyatt, Sarah (Inventor); Tsou, Pei-Lan (Inventor); Robertson, Dominique (Inventor); Boss, Wendy (Inventor)

2004-01-01

356

Ethics and Transgenic Crops: a Review  

Scientific Electronic Library Online (English)

Full Text Available SciELO Chile | Language: English Abstract in english This article represents a review of some of the ethical dilemmas that have arisen as a result of the development and deployment of transgenic crop plants. The potential for transgenic crops to alleviate human hunger and the possible effects on human health are discussed. Risks and benefits to the en [...] vironment resulting from genetic engineering of crops for resistance to biotic and abiotic stresses are considered, in addition to effects on biodiversity. The socio-economic impacts and distribution of benefits from transgenic technologies are reviewed. Fundamental issues of man’s relationship with nature and the environment, and theological matters are also addressed. An almost unprecedented amount of discussion has been stimulated on the merits and demerits of genetic engineering of crop plants, and has divided both the public and scientific communities. The arguments for and against transgenics are invariably based on visions of the new technology from widely different ethical perspectives.

Jonathan, Robinson.

1999-08-15

357

Ethical Issues in Genetic Engineering and Transgenics  

Science.gov (United States)

The issue-focused, peer-reviewed article demonstrates how transgenic technology has the potential of medical therapy, but it raises questions about these issues: creation of new life forms and crossing species boundaries, long-term effects on human health and the environment, blending of nonhuman animal and human DNA , and unintended personal, social, and cultural consequences.

Linda MacDonald Glenn (;)

2004-06-01

358

Transgenic Animals: Their Benefits To Human Welfare  

Science.gov (United States)

The issue-focused, reviewed, student article is about how transgenic animals, i.e., engineered to carry genes from other species, have the potential to improve human welfare in: agriculture, such as larger sheep that grow more wool, medicine, such as cows that produce insulin in their milk, andindustry, such as goats that produce spider silk for materials production.

Endang Tri Margawati (Bogor Agricultural University, Indonesia;)

2003-01-01

359

Metal resistance sequences and transgenic plants  

Science.gov (United States)

The present invention provides nucleic acid sequences encoding a metal ion resistance protein, which are expressible in plant cells. The metal resistance protein provides for the enzymatic reduction of metal ions including but not limited to divalent Cu, divalent mercury, trivalent gold, divalent cadmium, lead ions and monovalent silver ions. Transgenic plants which express these coding sequences exhibit increased resistance to metal ions in the environment as compared with plants which have not been so genetically modified. Transgenic plants with improved resistance to organometals including alkylmercury compounds, among others, are provided by the further inclusion of plant-expressible organometal lyase coding sequences, as specifically exemplified by the plant-expressible merB coding sequence. Furthermore, these transgenic plants which have been genetically modified to express the metal resistance coding sequences of the present invention can participate in the bioremediation of metal contamination via the enzymatic reduction of metal ions. Transgenic plants resistant to organometals can further mediate remediation of organic metal compounds, for example, alkylmetal compounds including but not limited to methyl mercury, methyl lead compounds, methyl cadmium and methyl arsenic compounds, in the environment by causing the freeing of mercuric or other metal ions and the reduction of the ionic mercury or other metal ions to the less toxic elemental mercury or other metals.

Meagher, Richard Brian (Athens, GA); Summers, Anne O. (Athens, GA); Rugh, Clayton L. (Athens, GA)

1999-10-12

360

Making BAC transgene constructs with lambda-red recombineering system for transgenic animals or cell lines.  

Science.gov (United States)

The genomic DNA libraries based on Bacteria Artificial Chromosomes (BAC) are the foundation of whole genomic mapping, sequencing, and annotation for many species like mice and humans. With their large insert size, BACs harbor the gene-of-interest and nearby transcriptional regulatory elements necessary to direct the expression of the gene-of-interest in a temporal and cell-type specific manner. When replacing a gene-of-interest with a transgene in vivo, the transgene can be expressed with the same patterns and machinery as that of the endogenous gene. This chapter describes in detail a method of using lambda-red recombineering to make BAC transgene constructs with the integration of a transgene into a designated location within a BAC. As the final BAC construct will be used for transfection in cell lines or making transgenic animals, specific considerations with BAC transgenes such as genotyping, BAC coverage and integrity as well as quality of BAC DNA will be addressed. Not only does this approach provide a practical and effective way to modify large DNA constructs, the same recombineering principles can apply to smaller high copy plasmids as well as to chromosome engineering. PMID:25239742

Holmes, Scott; Lyman, Suzanne; Hsu, Jen-Kang; Cheng, JrGang

2015-01-01

 
 
 
 
361

Generation of genetically modified rats from embryonic stem cells  

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At present, genetically modified rats have not been generated from ES cells because stable ES cells and a suitable injection method are not available. To monitor the pluripotency of rat ES cells, we generated Oct4-Venus transgenic (Tg) rats via a conventional method, in which Venus is expressed by the Oct4 promoter/enhancer. This monitoring system enabled us to define a significant condition of culture to establish authentic rat ES cells based on a combination of 20% FBS and cell signaling in...

Kawamata, Masaki; Ochiya, Takahiro

2010-01-01

362

Transgenic pig carrying green fluorescent proteasomes  

Science.gov (United States)

Among its many functions, the ubiquitin–proteasome system regulates substrate-specific proteolysis during the cell cycle, apoptosis, and fertilization and in pathologies such as Alzheimer’s disease, cancer, and liver cirrhosis. Proteasomes are present in human and boar spermatozoa, but little is known about the interactions of proteasomal subunits with other sperm proteins or structures. We have created a transgenic boar with green fluorescent protein (GFP) tagged 20S proteasomal core subunit ?-type 1 (PSMA1-GFP), hypothesizing that the PSMA1-GFP fusion protein will be incorporated into functional sperm proteasomes. Using direct epifluorescence imaging and indirect immunofluorescence detection, we have confirmed the presence of PSMA1-GFP in the sperm acrosome. Western blotting revealed a protein band corresponding to the predicted mass of PSMA1-GFP fusion protein (57 kDa) in transgenic spermatozoa. Transgenic boar fertility was confirmed by in vitro fertilization, resulting in transgenic blastocysts, and by mating, resulting in healthy transgenic offspring. Immunoprecipitation and proteomic analysis revealed that PSMA1-GFP copurifies with several acrosomal membrane-associated proteins (e.g., lactadherin/milk fat globule E8 and spermadhesin alanine-tryptophan-asparagine). The interaction of MFGE8 with PSMA1-GFP was confirmed through cross-immunoprecipitation. The identified proteasome-interacting proteins may regulate sperm proteasomal activity during fertilization or may be the substrates of proteasomal proteolysis during fertilization. Proteomic analysis also confirmed the interaction/coimmunoprecipitation of PSMA1-GFP with 13/14 proteasomal core subunits. These results demonstrate that the PSMA1-GFP was incorporated in the assembled sperm proteasomes. This mammal carrying green fluorescent proteasomes will be useful for studies of fertilization and wherever the ubiquitin–proteasome system plays a role in cellular function or pathology. PMID:23550158

Miles, Edward L.; O’Gorman, Chad; Zhao, Jianguo; Samuel, Melissa; Walters, Eric; Yi, Young-Joo; Prather, Randall S.; Wells, Kevin D.; Sutovsky, Peter

2013-01-01

363

Glucagon gene 5'-flanking sequences direct expression of simian virus 40 large T antigen to the intestine, producing carcinoma of the large bowel in transgenic mice.  

Science.gov (United States)

Glucagon and the glucagon-like peptides play important roles in the regulation of glucose homeostasis. Previous studies have demonstrated that approximately 1300 base pairs of rat glucagon gene 5'-flanking sequences direct transgene expression to the pancreas and brain, but not to the intestine, of transgenic mice. These observations suggested that different tissue-specific enhancer elements mediate activation of glucagon gene transcription in the pancreas and intestine. We have now generated mice that express SV40 large T antigen under the control of approximately 2000 base pairs of glucagon gene 5'-flanking sequences. Transgene expression was observed in the brain and pancreas in association with the development of pancreatic endocrine tumors. In contrast to the mice described previously, we also detected transgene expression throughout the gastrointestinal tract in endocrine cells of the stomach and small and large intestine. Focal areas of enteroendocrine cell hyperplasia in the large bowel invariably progressed to invasive and metastasizing plurihormonal endocrine carcinoma, which was clinically and pathologically evident by 4 weeks of age. In contrast, transgene expression in the small bowel and stomach was not associated with progression to either hyperplasia or carcinoma. The results of these studies provide functional evidence for the existence of an upstream cis-acting regulatory domain that directs glucagon gene transcription to the endocrine cells of the intestine in transgenic mice. PMID:1587847

Lee, Y C; Asa, S L; Drucker, D J

1992-05-25

364

Identification of Rat Rosa26 Locus Enables Generation of Knock-In Rat Lines Ubiquitously Expressing tdTomato  

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Recent discovery of a method for derivation and culture of germline-competent rat pluripotent stem cells (PSCs) enables generation of transgenic rats or knock-out rats via genetic modification of such PSCs. This opens the way to use rats, as is routine in mice, for analyses of gene functions or physiological features. In mouse or human, one widely used technique to express a gene of interest stably and ubiquitously is to insert that gene into the Rosa26 locus via gene targeting of PSCs. Rosa2...

Kobayashi, Toshihiro; Kato-itoh, Megumi; Yamaguchi, Tomoyuki; Tamura, Chihiro; Sanbo, Makoto; Hirabayashi, Masumi; Nakauchi, Hiromitsu

2012-01-01

365

Green Tea Polyphenols Control Dysregulated Glutamate Dehydrogenase in Transgenic Mice by Hijacking the ADP Activation Site  

Energy Technology Data Exchange (ETDEWEB)

Glutamate dehydrogenase (GDH) catalyzes the oxidative deamination of L-glutamate and, in animals, is extensively regulated by a number of metabolites. Gain of function mutations in GDH that abrogate GTP inhibition cause the hyperinsulinism/hyperammonemia syndrome (HHS), resulting in increased pancreatic {beta}-cell responsiveness to leucine and susceptibility to hypoglycemia following high protein meals. We have previously shown that two of the polyphenols from green tea (epigallocatechin gallate (EGCG) and epicatechin gallate (ECG)) inhibit GDH in vitro and that EGCG blocks GDH-mediated insulin secretion in wild type rat islets. Using structural and site-directed mutagenesis studies, we demonstrate that ECG binds to the same site as the allosteric regulator, ADP. Perifusion assays using pancreatic islets from transgenic mice expressing a human HHS form of GDH demonstrate that the hyperresponse to glutamine caused by dysregulated GDH is blocked by the addition of EGCG. As observed in HHS patients, these transgenic mice are hypersensitive to amino acid feeding, and this is abrogated by oral administration of EGCG prior to challenge. Finally, the low basal blood glucose level in the HHS mouse model is improved upon chronic administration of EGCG. These results suggest that this common natural product or some derivative thereof may prove useful in controlling this genetic disorder. Of broader clinical implication is that other groups have shown that restriction of glutamine catabolism via these GDH inhibitors can be useful in treating various tumors. This HHS transgenic mouse model offers a highly useful means to test these agents in vivo.

Li, Changhong; Li, Ming; Chen, Pan; Narayan, Srinivas; Matschinsky, Franz M.; Bennett, Michael J.; Stanley, Charles A.; Smith, Thomas J. (CH-PA); (UPENN); (Danforth)

2012-05-09

366

The distribution of transgene insertion sites in barley determined by physical and genetic mapping.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The exact site of transgene insertion into a plant host genome is one feature of the genetic transformation process that cannot, at present, be controlled and is often poorly understood. The site of transgene insertion may have implications for transgene stability and for potential unintended effects of the transgene on plant metabolism. To increase our understanding of transgene insertion sites in barley, a detailed analysis of transgene integration in independently derived transgenic barley...

Salvo-garrido, Haroldo; Travella, Silvia; Bilham, Lorelei J.; Harwood, Wendy A.; Snape, John W.

2004-01-01

367

Assessment of peanut quality and compositional characteristics among transgenic sclerotinia blight-resistant and non-transgenic susceptible cultivars.  

Science.gov (United States)

This study presents the results of a comparison that includes an analysis of variance and a canonical discriminant analysis to determine compositional equivalence and similarity between transgenic, sclerotinia blight-resistant and non-transgenic, susceptible cultivars of peanut in 3 years of field trials. Three Virginia-type cultivars (NC 7, Wilson, and Perry) and their corresponding transgenic lines (N70, W73, and P39) with a barley oxalate oxidase gene were analyzed for differences in key mineral nutrients, fatty acid components, hay constituents, and grade characteristics. Results from both analyses demonstrated that transgenic lines were compositionally similar to their non-transgenic parent cultivar in all factors as well as market-grade characteristics and nutritional value. Transgenic lines expressing oxalate oxidase for resistance to sclerotinia blight were substantially equivalent to their non-transgenic parent cultivar in quality and compositional characteristics. PMID:24972023

Hu, Jiahuai; Telenko, Darcy E P; Phipps, Patrick M; Grabau, Elizabeth A

2014-08-01

368

Wading pools and fading memories – place navigation in transgenic mouse models of Alzheimer’s disease  

Directory of Open Access Journals (Sweden)

Full Text Available The Morris swim navigation task (‘water maze’ has been a primary research tools to assess hippocampal delpendent spatial learning and memory is rodents for three decades. Originally developed for rats, its application to mouse studies has been a tedious process, but nowadays there are more studies performed with the Morris swim task in mice than in rats. The task has proved to be particularly useful in demonstrating age-related memory impairment in transgenic mouse models of Alzheimer’s disease. This review focuses on task details that are most relevant for its application to mouse studies in general and characteristic patterns of impaired performance in Alzheimer model mice as compared with rodents sustaining hippocampal lesions.

HeikkiTanila

2012-06-01

369

Generation of genetically modified rats from embryonic stem cells.  

Science.gov (United States)

At present, genetically modified rats have not been generated from ES cells because stable ES cells and a suitable injection method are not available. To monitor the pluripotency of rat ES cells, we generated Oct4-Venus transgenic (Tg) rats via a conventional method, in which Venus is expressed by the Oct4 promoter/enhancer. This monitoring system enabled us to define a significant condition of culture to establish authentic rat ES cells based on a combination of 20% FBS and cell signaling inhibitors for Rho-associated kinase, mitogen-activated protein kinase, TGF-beta, and glycogen synthase kinase-3. The rat ES cells expressed ES cell markers such as Oct4, Nanog, Sox2, and Rex1 and retained a normal karyotype. Embryoid bodies and teratomas were also produced from the rat ES cells. All six ES cell lines derived from three different rat strains successfully achieved germline transmission, which strongly depended on the presence of the inhibitors during the injection process. Most importantly, high-quality Tg rats possessing a correct transgene expression pattern were successfully generated via the selection of gene-manipulated ES cell clones through germline transmission. Our rat ES cells should be sufficiently able to receive gene targeting as well as Tg manipulation, thus providing valuable animal models for the study of human diseases. PMID:20660726

Kawamata, Masaki; Ochiya, Takahiro

2010-08-10

370

Compensation of the AKT signaling by ERK signaling in transgenic mice hearts overexpressing TRIM72  

Energy Technology Data Exchange (ETDEWEB)

The AKT and ERK signaling pathways are known to be involved in cell hypertrophy, proliferation, survival and differentiation. Although there is evidence for crosstalk between these two signaling pathways in cellulo, there is less evidence for cross talk in vivo. Here, we show that crosstalk between AKT and ERK signaling in the hearts of TRIM72-overexpressing transgenic mice (TRIM72-Tg) with alpha-MHC promoter regulates and maintains their heart size. TRIM72, a heart- and skeletal muscle-specific protein, downregulates AKT-mTOR signaling via IRS-1 degradation and reduces the size of rat cardiomyocytes and the size of postnatal TRIM72-Tg hearts. TRIM72 expression was upregulated by hypertrophic inducers in cardiomyocytes, while IRS-1 was downregulated by IGF-1. TRIM72 specifically regulated IGF-1-dependent AKT-mTOR signaling, resulting in a reduction of the size of cardiomyocytes. Postnatal TRIM72-Tg hearts were smaller than control-treated hearts with inhibition of AKT-mTOR signaling. However, adult TRIM72-Tg hearts were larger than of control despite the suppression of AKT-mTOR signaling. Activation of ERK, PKC-?, and JNK were observed to be elevated in adult TRIM72-Tg, and these signals were mediated by ET-1 via the ET receptors A and B. Altogether, these results suggest that AKT signaling regulates cardiac hypertrophy in physiological conditions, and ERK signaling compensates for the absence of AKT signaling during TRIM72 overexpression, leading to pathological hypertrophy. -- Highlights: • TRIM72 inhibits AKT signaling through ubiquitination of IRS-1 in cardiac cells. • TRIM72 regulates the size of cardiac cells. • TRIM72 regulates size of postnatal TRIM72-overexpressing transgenic mice hearts. • Adult TRIM72-overexpressing transgenic mice hearts showed cardiac dysfunction. • Adult TRIM72 transgenic mice hearts showed higher expression of endothelin receptors.

Ham, Young-Mi, E-mail: youngmi_ham@hms.harvard.edu [College of Life Science and Biotechnology, Korea University, Seoul (Korea, Republic of); Department of Cell Biology, Harvard Medical School, Boston, MA 02115 (United States); Mahoney, Sarah Jane [Department of Cell Biology, Harvard Medical School, Boston, MA 02115 (United States)

2013-06-10

371

Compensation of the AKT signaling by ERK signaling in transgenic mice hearts overexpressing TRIM72  

International Nuclear Information System (INIS)

The AKT and ERK signaling pathways are known to be involved in cell hypertrophy, proliferation, survival and differentiation. Although there is evidence for crosstalk between these two signaling pathways in cellulo, there is less evidence for cross talk in vivo. Here, we show that crosstalk between AKT and ERK signaling in the hearts of TRIM72-overexpressing transgenic mice (TRIM72-Tg) with alpha-MHC promoter regulates and maintains their heart size. TRIM72, a heart- and skeletal muscle-specific protein, downregulates AKT-mTOR signaling via IRS-1 degradation and reduces the size of rat cardiomyocytes and the size of postnatal TRIM72-Tg hearts. TRIM72 expression was upregulated by hypertrophic inducers in cardiomyocytes, while IRS-1 was downregulated by IGF-1. TRIM72 specifically regulated IGF-1-dependent AKT-mTOR signaling, resulting in a reduction of the size of cardiomyocytes. Postnatal TRIM72-Tg hearts were smaller than control-treated hearts with inhibition of AKT-mTOR signaling. However, adult TRIM72-Tg hearts were larger than of control despite the suppression of AKT-mTOR signaling. Activation of ERK, PKC-?, and JNK were observed to be elevated in adult TRIM72-Tg, and these signals were mediated by ET-1 via the ET receptors A and B. Altogether, these results suggest that AKT signaling regulates cardiac hypertrophy in physiological conditions, and ERK signaling compensates for the absence of AKT signaling during TRIM72 overexpression, leading to pathological hypertrophy. -- Highlights: • TRIM72 inhibits AKT signaling through ubiquitination of IRS-1 in cardiac cells. • TRIM72 regulates the size of cardiac cells. • TRIM72 regulates size of postnatal TRIM72-overexpressing transgenic mice hearts. • Adult TRIM72-overexpressing transgenic mice hearts showed cardiac dysfunction. • Adult TRIM72 transgenic mice hearts showed higher expression of endothelin receptors

372

A multidisciplinary approach involving comparative 'OMICS' of transgenic and non-transgenic soybeam seeds  

International Nuclear Information System (INIS)

Complete text of publication follows. Soybean culture has an expressive impact in the economy of many countries, being the commercialization of its by-products, which presents many benefits in terms of health and nutritional aspects, and which also includes a fuel alternative (biodiesel), the main factor for the large soybean production. Part of this impact is due to the transgenic modification of soybean, conferring enhanced characteristics to the culture, such as tolerance to fungicides (Y. Kim et al., J. Microbiol. Biotechnol., 16 (2006), 25-31.). Due to the insertion of hexogen genes, some proteome modification is possible (S. Natarajan et al., Anal. Biochem., 342 (2005), 214-220), and recently some metallome modification was reported by our research group (A. Sussulini et al., J. Anal. At. Spectrom., 22 (2007), 1501-1506). Then, the aim of this work is to enlarge the results in terms of 'omics' when considering transgenic and non-transgenic soybean seeds. For this task, the identification of more than 140 soybean proteins using MALDI-QTOF-MS after 2D-PAGE protein separation (369±46 and 376±42 protein spots in the 4-7 pI range for transgenic and non-transgenic soybean seeds, respectively), the analysis of the protein expression using image program, the analysis of some enzymes (SOD, GR, APX, CAT) involved in the ROS production, the mapping of 80 protein spots using SR-XRF, and the metal identification of more than 30 spots using ICP-MS was carried out. In terms using ICP-MS was carried out. In terms of metal distribution when considering some proteins, the results displayed a great ability of proteins bind different metal ions. High iron (sucrose binding protein homolog S-64 - 57,922 kDa), chromium (protein not identified), lead (seed maturation protein PM 41 - 15,103 kDa), copper and tin (trypsin inhibitor (kunitz), chain A - 20,417 kDa) contents were achieved in the non-transgenic soybean, while high magnesium (actin - 50,281 kDa), barium (protein not identified) and ruthenium (protein not identified) contents were achieved in the transgenic soybean. The results put in evidence the possibility to find a biomarker candidate for differentiating transgenic and non-transgenic organisms. Financial support from Fundacao de Amparo a Pesquisa do Estado de Sao Paulo - FAPESP, Conselho Nacional de Desenvolvimento Cientifico e Tecnologico - CNPq, Fianciadora de Estudos e Projetos - FINEP, and Proteome Network of the Sao Paulo state - Brazilian National Laboratory of Synchrotron Radiation are highly acknowledged.

373

Effect of transgene number of spontaneous and radiation-induced micronuclei in lacl transgenic mice  

International Nuclear Information System (INIS)

Lacl transgenic mice are widely used for the measurement of mutations in specific target issues. The lacl transgene is present in mice as 40 tandem repeats; this sequence is homozygous (contained in both copies of chromosome 5) in C57Bl/6 mice, and is hemizygous in B6C3F1 mice. Previous reports have indicated that tandem repeats can produce chromosome instability, fragile sites, and other effects. To determine whether the presence of the transgene effects micronucleus induction we compared the response of nontransgenic (NTR) to hemizygous (HEMI) transgenic B6C3F1 mice and to hemizygous and homozygous (HOMO) transgenic C57Bl/6 mice. Five mice/group were irradiated with 500 cGy from a 137Cs source. Bone marrow was harvested 24 hr after treatment and 2000 polychromatic erythrocytes (PCE) were analyzed per animal. The presence or absence of the lacl transgene had no effect in unirradiated mice on the percent of micronucleated PCE (MN) or on the ratio of PCE to total red blood cells for either strain: B6C3F1 mice had MN frequencies of 0.26% and 0.20% for NTR and HEMI mice, respectively; C57Bl/6 mice had MN frequencies of 0.34%, 0.32%, and 0.38% for NTR, HEMI, and HOMO mice, respectively. Radiation-induced micronucleus frequencies were significantly higher in HEMI lacl B6C3F1 mice (2.85%) than in NTR litter mates (1.59%); the converse was true in C57Bl/6 mice: NTR were 2.45%, HEMI were 1.25%, HOMO were 1.65%. These data suggest that the lacl transgene does not cauggest that the lacl transgene does not cause chromosome instability as measured by spontaneous micronucleus levels. However, the response of these transgenic mice to a variety of clastogenic agents needs to be investigated before they are integrated into standard in vivo assays for chromosome damage

374

Transgenic arthropods and the sterile insect technique  

International Nuclear Information System (INIS)

The Sterile Insect Technique can benefit from transgenesis in three ways by creating; (1) genetically marked strains, (2) genetic sexing strains and (3) strains that induce molecular sterility in the field. Experience with the development of genetic sexing strains based on indicates that caution is required during the experimental evaluation of any potential transgenic strain. Two major scientific concerns involve the overall fitness of transgenic strains and their stability over time. The latter being very important especially when the extremely large numbers of insects that are mass reared is taken into account. Currently transformation events are random and it will probably be necessary to select suitable strains from many that are induced. The success of transformation itself in many insect species will enable many new strategies to be developed and tested. (author)

375

Transgenic nonhuman primates for neurodegenerative diseases  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Animal models that represent human diseases constitute an important tool in understanding the pathogenesis of the diseases, and in developing effective therapies. Neurodegenerative diseases are complex disorders involving neuropathologic and psychiatric alterations. Although transgenic and knock-in mouse models of Alzheimer's disease, (AD, Parkinson's disease (PD and Huntington's disease (HD have been created, limited representation in clinical aspects has been recognized and the rodent models lack true neurodegeneration. Chemical induction of HD and PD in nonhuman primates (NHP has been reported, however, the role of intrinsic genetic factors in the development of the diseases is indeterminable. Nonhuman primates closely parallel humans with regard to genetic, neuroanatomic, and cognitive/behavioral characteristics. Accordingly, the development of NHP models for neurodegenerative diseases holds greater promise for success in the discovery of diagnoses, treatments, and cures than approaches using other animal species. Therefore, a transgenic NHP carrying a mutant gene similar to that of patients will help to clarify our understanding of disease onset and progression. Additionally, monitoring disease onset and development in the transgenic NHP by high resolution brain imaging technology such as MRI, and behavioral and cognitive testing can all be carried out simultaneously in the NHP but not in other animal models. Moreover, because of the similarity in motor repertoire between NHPs and humans, it will also be possible to compare the neurologic syndrome observed in the NHP model to that in patients. Understanding the correlation between genetic defects and physiologic changes (e.g. oxidative damage will lead to a better understanding of disease progression and the development of patient treatments, medications and preventive approaches for high risk individuals. The impact of the transgenic NHP model in understanding the role which genetic disorders play in the development of efficacious interventions and medications is foreseeable.

Chan Anthony WS

2004-06-01

376

Use of new transgenic mouse models  

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The use of transgenic mouse models with green fluorescent astrocytes and red fluorescent oligodendrocytes allows the direct visualisation of cellular responses in the healthy and injured brain. After acute traumatic lesions or ischemic stroke in the neocortex fast neuronal and glial cell death is frequently observed. Activations of different glial cells like astroglia, oligodendrocytes and microglia cells are sequelae of secondary injury processes. A complex and not yet underst...

Scheller, Anja

2010-01-01

377

A neuroglobin-overexpressing transgenic mouse  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Neuroglobin (Ngb) is a recently discovered vertebrate globin expressed primarily in neurons. Ngb expression is induced by hypoxia and ischemia, and Ngb protects neurons from these insults. However, its normal physiological role and the mechanism underlying its neuroprotective action are uncertain. We report production of a transgenic mouse in which Ngb is overexpressed under the control of the chicken ?-actin promoter. This mouse should prove helpful for studying Ngb-mediated effects in vitr...

Khan, Adil A.; Sun, Yunjuan; Jin, Kunlin; Mao, Xiao Ou; Chen, Sylvia; Ellerby, Lisa M.; Greenberg, David A.

2007-01-01

378

Overview: Engineering transgenic constructs and mice  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Cell biology research encompasses everything from single cells to whole animals. Recent discoveries concerning particular gene functions can be applied to the whole animal for understanding genotype-phenotype relationships underlying disease mechanisms. For this reason, genetically manipulated mouse models are now considered essential to correctly understand disease processes in whole animals. This unit provides the basic mouse technologies used to generate conventional transgenic mice, which...

Haruyama, Naoto; Cho, Andrew; Kulkarni, Ashok B.

2009-01-01

379

T-antigen transgenic mouse models  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The study of polyomavirus has benefited immensely from two scientific methodologies, cell culture and in vitro studies on one side and the use of transgenic mice as experimental models on the other. Both approaches allowed us to identify cellular products targeted by the viruses, the consequences of these interactions at the phenotypic and molecular level, and thus the potential roles of the targets within their normal cellular context. In particular, cell culture and in vitro reports suggest...

Robles, Maria Teresa Sa?enz; Pipas, James M.

2009-01-01

380

Analysis of somatic mutations in kappa transgenes  

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We have examined the nature and localization of somatic mutations in three kappa transgenes cloned from IgG-secreting hybridomas. All of the mutations identified were single base substitutions. Mutations were localized to the variable (V) region and its flanking sequences. In every case, the nuclear matrix association region, kappa enhancer, and C gene were spared. These data indicate that the rearranged kappa gene contains the necessary sequences for targeting of the mutation process, and su...

1990-01-01

 
 
 
 
381

Transgenic pig carrying green fluorescent proteasomes  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Among its many functions, the ubiquitin–proteasome system regulates substrate-specific proteolysis during the cell cycle, apoptosis, and fertilization and in pathologies such as Alzheimer’s disease, cancer, and liver cirrhosis. Proteasomes are present in human and boar spermatozoa, but little is known about the interactions of proteasomal subunits with other sperm proteins or structures. We have created a transgenic boar with green fluorescent protein (GFP) tagged 20S proteasomal core sub...

Miles, Edward L.; O’gorman, Chad; Zhao, Jianguo; Samuel, Melissa; Walters, Eric; Yi, Young-joo; Sutovsky, Miriam; Prather, Randall S.; Wells, Kevin D.; Sutovsky, Peter

2013-01-01

382

Zebrafish transgenic Enhancer TRAP line database (ZETRAP)  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Abstract Background The zebrafish, Danio rerio, is used as a model organism to study vertebrate genetics and development. An effective enhancer trap (ET) in zebrafish using the Tol2 transposon has been demonstrated. This approach could be used to study embryogenesis of a vertebrate species in real time and with high resolution. Description The information gathered during the course of systematic investigation of many ET transgenic lines have be...

Emelyanov Alexander; Parinov Sergey; Kondrichin Igor; Gh, Choo Benjamin; Go William; Toh Wei-chang; Korzh Vladimir

2006-01-01

383

Biological containment strategies for transgenic crops  

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Biological containment is the prevention or reduction in the spread of transgenes by modifying plant growth or development, most commonly through modification of reproductive characteristics. This review provides a summary of the current strategies for biological containment, including the use of both naturally occurring and engineered mechanisms. A wide range of strategies and their efficacy, where possible, are discussed. While some strategies are still in the conceptual phase, others are m...

Maagd, R. A.; Boutilier, K. A.

2013-01-01

384

Regulation of transgene expression in genetic immunization  

Directory of Open Access Journals (Sweden)

Full Text Available The use of mammalian gene expression vectors has become increasingly important for genetic immunization and gene therapy as well as basic research. Essential for the success of these vectors in genetic immunization is the proper choice of a promoter linked to the antigen of interest. Many genetic immunization vectors use promoter elements from pathogenic viruses including SV40 and CMV. Lymphokines produced by the immune response to proteins expressed by these vectors could inhibit further transcription initiation by viral promoters. Our objective was to determine the effect of IFN-g on transgene expression driven by viral SV40 or CMV promoter/enhancer and the mammalian promoter/enhancer for the major histocompatibility complex class I (MHC I gene. We transfected the luciferase gene driven by these three promoters into 14 cell lines of many tissues and several species. Luciferase assays of transfected cells untreated or treated with IFN-g indicated that although the viral promoters could drive luciferase production in all cell lines tested to higher or lower levels than the MHC I promoter, treatment with IFN-g inhibited transgene expression in most of the cell lines and amplification of the MHC I promoter-driven transgene expression in all cell lines. These data indicate that the SV40 and CMV promoter/enhancers may not be a suitable choice for gene delivery especially for genetic immunization or cancer cytokine gene therapy. The MHC I promoter/enhancer, on the other hand, may be an ideal transgene promoter for applications involving the immune system.

J.S. Harms

1999-02-01

385

Regulation of transgene expression in genetic immunization  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english The use of mammalian gene expression vectors has become increasingly important for genetic immunization and gene therapy as well as basic research. Essential for the success of these vectors in genetic immunization is the proper choice of a promoter linked to the antigen of interest. Many genetic im [...] munization vectors use promoter elements from pathogenic viruses including SV40 and CMV. Lymphokines produced by the immune response to proteins expressed by these vectors could inhibit further transcription initiation by viral promoters. Our objective was to determine the effect of IFN-g on transgene expression driven by viral SV40 or CMV promoter/enhancer and the mammalian promoter/enhancer for the major histocompatibility complex class I (MHC I) gene. We transfected the luciferase gene driven by these three promoters into 14 cell lines of many tissues and several species. Luciferase assays of transfected cells untreated or treated with IFN-g indicated that although the viral promoters could drive luciferase production in all cell lines tested to higher or lower levels than the MHC I promoter, treatment with IFN-g inhibited transgene expression in most of the cell lines and amplification of the MHC I promoter-driven transgene expression in all cell lines. These data indicate that the SV40 and CMV promoter/enhancers may not be a suitable choice for gene delivery especially for genetic immunization or cancer cytokine gene therapy. The MHC I promoter/enhancer, on the other hand, may be an ideal transgene promoter for applications involving the immune system.

J.S., Harms; S.C., Oliveira; G.A., Splitter.

1999-02-01

386

Transgene Detection by Digital Droplet PCR  

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Somatic gene therapy is a promising tool for the treatment of severe diseases. Because of its abuse potential for performance enhancement in sports, the World Anti-Doping Agency (WADA) included the term ‘gene doping’ in the official list of banned substances and methods in 2004. Several nested PCR or qPCR-based strategies have been proposed that aim at detecting long-term presence of transgene in blood, but these strategies are hampered by technical limitations. We developed a digital dro...

Moser, Dirk A.; Braga, Luca; Raso, Andrea; Zacchigna, Serena; Giacca, Mauro; Simon, Perikles

2014-01-01

387

Toxins for Transgenic Resistance to Hemipteran Pests  

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The sap sucking insects (Hemiptera), which include aphids, whiteflies, plant bugs and stink bugs, have emerged as major agricultural pests. The Hemiptera cause direct damage by feeding on crops, and in some cases indirect damage by transmission of plant viruses. Current management relies almost exclusively on application of classical chemical insecticides. While the development of transgenic crops expressing toxins derived from the bacterium Bacillus thuringiensis (Bt) ha...

Bonning, Bryony C.; Chougule, Nanasaheb P.

2012-01-01

388

Selective expression of trypsin fusion genes in acinar cells of the pancreas and stomach of transgenic mice.  

Science.gov (United States)

Fusion genes combining the 5'-transcriptional regulatory region of the rat trypsin I gene and the structural gene of human growth hormone as a reporter were expressed to the high levels characteristic of the endogenous trypsin I gene selectively in the acinar cells of the pancreas of transgenic mice. As little as 232 base pairs of trypsin gene sequences containing the transcriptional start site and upstream promoter elements were sufficient to direct pancreatic expression. The tissue-specific expression was controlled transcriptionally. Trypsin-human growth hormone fusion transgenes also were expressed, although at low levels, in the stomach, an unexpected site for the expression of pancreatic digestive enzymes. Expression in the stomach of endogenous trypsin, elastase, and amylase genes in both normal and transgenic mice verified that transgene expression was consistent with normal expression of pancreatic genes. Endogenous amylase colocalizes with pepsinogen in the acinar cell-like Chief cells of the glandular portion of the mouse stomach. The expression of pancreatic genes in stomach cells is probably the consequence of similar developmental origins of pancreatic and gastric acinar cells from the primordial gut. PMID:1464618

Davis, B P; Hammer, R E; Messing, A; MacDonald, R J

1992-12-25

389

Dual reporter transgene driven by 2.3Col1a1 promoter is active in differentiated osteoblasts  

Science.gov (United States)

AIM: As quantitative and spatial analyses of promoter reporter constructs are not easily performed in intact bone, we designed a reporter gene specific to bone, which could be analyzed both visually and quantitatively by using chloramphenicol acetyltransferase (CAT) and a cyan version of green fluorescent protein (GFPcyan), driven by a 2.3-kb fragment of the rat collagen promoter (Col2.3). METHODS: The construct Col2.3CATiresGFPcyan was used for generating transgenic mice. Quantitative measurement of promoter activity was performed by CAT analysis of different tissues derived from transgenic animals; localization was performed by visualized GFP in frozen bone sections. To assess transgene expression during in vitro differentiation, marrow stromal cell and neonatal calvarial osteoblast cultures were analyzed for CAT and GFP activity. RESULTS: In mice, CAT activity was detected in the calvaria, long bone, teeth, and tendon, whereas histology showed that GFP expression was limited to osteoblasts and osteocytes. In cell culture, increased activity of CAT correlated with increased differentiation, and GFP activity was restricted to mineralized nodules. CONCLUSION: The concept of a dual reporter allows a simultaneous visual and quantitative analysis of transgene activity in bone.

Marijanovic, Inga; Jiang, Xi; Kronenberg, Mark S.; Stover, Mary Louise; Erceg, Ivana; Lichtler, Alexander C.; Rowe, David W.

2003-01-01

390

Arsenic biotransformation and volatilization in transgenic rice.  

Science.gov (United States)

• Biotransformation of arsenic includes oxidation, reduction, methylation, and conversion to more complex organic arsenicals. Members of the class of arsenite (As(III)) S-adenosylmethyltransferase enzymes catalyze As(III) methylation to a variety of mono-, di-, and trimethylated species, some of which are less toxic than As(III) itself. However, no methyltransferase gene has been identified in plants. • Here, an arsM gene from the soil bacterium Rhodopseudomonas palustris was expressed in Japonica rice (Oryza sativa) cv Nipponbare, and the transgenic rice produced methylated arsenic species, which were measured by inductively coupled plasma mass spectrometry (ICP-MS) and high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP-MS). • Both monomethylarsenate (MAs(V)) and dimethylarsenate (DMAs(V)) were detected in the roots and shoots of transgenic rice. After 12 d exposure to As(III), the transgenic rice gave off 10-fold greater volatile arsenicals. • The present study demonstrates that expression of an arsM gene in rice induces arsenic methylation and volatilization, theoretically providing a potential stratagem for phytoremediation. PMID:21517874

Meng, Xiang-Yan; Qin, Jie; Wang, Li-Hong; Duan, Gui-Lan; Sun, Guo-Xin; Wu, Hui-Lan; Chu, Cheng-Cai; Ling, Hong-Qing; Rosen, Barry P; Zhu, Yong-Guan

2011-07-01

391

Transgenic approaches to western corn rootworm control.  

Science.gov (United States)

The western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) is a significant corn pest throughout the United States corn belt. Rootworm larvae feed on corn roots causing yield losses and control expenditures that are estimated to exceed US$1 billion annually. Traditional management practices to control rootworms such as chemical insecticides or crop rotation have suffered reduced effectiveness due to the development of physiological and behavioral resistance. Transgenic maize expressing insecticidal proteins are very successful in protecting against rootworm damage and preserving corn yield potential. However, the high rate of grower adoption and early reliance on hybrids expressing a single mode of action and low-dose traits threatens the durability of commercialized transgenic rootworm technology for rootworm control. A summary of current transgenic approaches for rootworm control and the corresponding insect resistance management practices is included. An overview of potential new modes of action based on insecticidal proteins, and especially RNAi targeting mRNA coding for essential insect proteins is provided. PMID:23604211

Narva, Kenneth E; Siegfried, Blair D; Storer, Nicholas P

2013-01-01

392

Comparing the Agronomic and Grain Quality Characteristics of Transgenic Rice Lines Expressing cry1Ab vs. Non-Transgenic Controls  

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Full Text Available This study aimed to investigate and compare the agronomic and grain quality attributes of three advanced backcross-derived transgenic rice lines expressing synthetic cry1Ab gene vs. non-transgenic control in a Randomized Complete Block Design (RCBD under field conditions. The data exhibited that transgenic rice lines, Neda and Nemat were higher in height, earlier in maturity and highly resistant to striped stem borer (Chilo suppressalis in comparing with non-transgenic varieties. In contrast, no significant difference was observed for transgenic Khazar as compared to its control, except for 1000-grain weight. Laboratory tests for grain physicochemical properties showed no significant variations between transgenic lines and non-transgenic controls. However, some variations for traits like Amylose Content (AC and Gel Consistency (GC were seen for transgenic Neda and Khazar, respectively. As regards the rice striped stem borer natural infestation, field-release experiment indicated that all three transgenic rice lines conferred a very high degree of resistance to rice striped stem borer as compared to non-transgenic check varieties.

G. Kiani

2009-01-01

393

Production of recombinant proteins in milk of transgenic and non-transgenic goats  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english Among all the transgenic mammalians produced so far, goats have represented an excellent model of transgenesis when considering the factors such as the market demand for protein, volume of milk produced per lactation and reproductive rate. Various recombinant proteins have been obtained from the tra [...] nsgenic and non-transgenic goats, and among these, human antithrombin, produced by the transgenic goats, was the first recombinant protein of animal origin to be released as a drug for the clinical use in humans. This review reports the aspects inherent to the production of recombinant proteins in the goats, from the production of the animal bioreactors up to the expression of these proteins in their milk.

Raylene Ramos, Moura; Luciana Magalhães, Melo; Vicente José de Figueirêdo, Freitas.

2011-10-01

394

Transgenic Rice Expressing Amyloid ?-peptide for Oral Immunization  

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Various vaccine therapies for Alzheimer's disease (AD) have been investigated. Here we report transgenic rice expressing amyloid ?-peptide (A?). The A?42 gene fused with a green fluorescent protein gene was introduced into rice using the Agrobacterium method. When transgenic brown rice expressing A? was orally administered to mice, serum anti-A? antibody titers were elevated. The same results were observed when mice were fed boiled, transgenic brown rice. Th...

Taiji Yoshida, Eiichi Kimura

2011-01-01

395

Generation of bovine transgenics using somatic cell nuclear transfer  

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Abstract The ability to produce transgenic animals through the introduction of exogenous DNA has existed for many years. However, past methods available to generate transgenic animals, such as pronuclear microinjection or the use of embryonic stem cells, have either been inefficient or not available in all animals, bovine included. More recently somatic cell nuclear transfer has provided a method to create transgenic animals that overcomes many deficiencies present in other methods....

Stice Steven L; Hodges Craig A

2003-01-01

396

Toxin–antitoxin based transgene expression in mammalian cells  

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Long-term, recombinant gene expression in mammalian cells depends on the nature of the transgene integration site and its inherent properties to modulate transcription (epigenetic effects). Here we describe a method by which high transgene expression is achieved and stabilized in extensively proliferating cultures. The method is based on strict co-expression of the transgene with an antitoxin in cells that express the respective toxin. Since the strength of antitoxin expression correlates wit...

Nehlsen, K.; Herrmann, S.; Zauers, J.; Hauser, H.; Wirth, D.

2010-01-01

397

Hepatic steatosis in transgenic mice overexpressing human histone deacetylase 1  

International Nuclear Information System (INIS)

It is generally thought that histone deacetylases (HDACs) play important roles in the transcriptional regulation of genes. However, little information is available concerning the specific functions of individual HDACs in disease states. In this study, two transgenic mice lines were established which harbored the human HDAC1 gene. Overexpressed HDAC1 was detected in the nuclei of transgenic liver cells, and HDAC1 enzymatic activity was significantly higher in the transgenic mice than in control littermates. The HDAC1 transgenic mice exhibited a high incidence of hepatic steatosis and nuclear pleomorphism. Molecular studies showed that HDAC1 may contribute to nuclear pleomorphism through the p53/p21 signaling pathway

398

Comparative metallomics of transgenic and non-transgenic soybeans using HPLC-ICP-MS  

International Nuclear Information System (INIS)

Complete text of publication follows. In the last years, many soybean varieties have been developed, and due to these modifications, the proteins composition and profile can be affected, causing changes in the species proteome (S. Natarajan et al., Anal. Biochem., 342 (2005), 214-220.). With the proteome modifications, the metallome of this specie, defined as the total content of metals and metalloids in a cell or tissue (J. Spuznar, Analyst, 130 (2005), 442-465.), can also be affected (A. Sussulini et al., J. Anal. At. Spectrom., 22 (2007), 1501-1506.). So, the aim of this work is to amplify the information about the transgenic and non-transgenic soybeans metallome, and doing that we expect to find biomarkers that can differentiate the transgenic and non-transgenic soybeans physiologically. For that purpose a SEC column (GE Healthcare, model Superdex 200) was employed for the separation of the proteins, which were extracted using the mobile phase of the chromatographic system (90 mmol.L-1 phosphate buffer - pH 7.2). After the chromatographic separation, the eluate was passed through a DAD Series 200 detector (PerkinElmer), the fractions were collected and latter introduced into the ICP-MS (PerkinElmer, model ELANDRC-e) for the element-selective detection. The calibration of the column using purified proteins of known molecular weight allowed the calculation of the approximate masses of the eight fractions (1800-800 kDa; 800-420 kDa; 420-120 kDa; 100-23 kDa kDa; 800-420 kDa; 420-120 kDa; 100-23 kDa; 23-7 kDa; 7-2 kDa; 2-0.4 kDa and 0.4-0.2 kDa, respectively) identified in the transgenic and non-transgenic soybeans after 95 min of separation using a flow rate of 0.25 mL.min-1. A wide range of elements could be identified in all the fractions, including: Cu, Zn, Mn, Mg, Ni, Cr, Hg, Fe and Pb. Differences in the detectability of elements in the transgenic and non-transgenic soybeans were found, specially for Hg where the counts were two times higher in the transgenic soybean. Elements were found in the two samples that were not common for both of them, such as Sr identified only in fraction 2 of the non-transgenic soybean and Th in fraction 4 of the transgenic soybean. Financial support from Fundacao de Amparo a Pesquisa do Estado de Sao Paulo - FAPESP and Conselho Nacional de Desenvolvimento Cientifico e Tecnologico - CNPq are highly acknowledged.

399

An analytical model assessing the potential threat to natural habitats from insect resistance transgenes: continuous transgene input  

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The potential effects of ‘escape’ of genetically modified material (transgenes) into natural communities is a major concern in their use. These effects may be limited in the first instance by limiting the proportion of transgene-carrying plants in the natural community. We previously presented an analytical model of the ecological processes governing the relative abundance and persistence of insect resistance (IR) transgenes in a natural community. In that paper, we illustrated the case i...

Kelly, Colleen K.; Bowler, Michael; Breden, Felix

2006-01-01

400

Rheumatic manifestations of inflammatory bowel disease  

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Full Text Available This article reviews the literature concerning rheumatic manifestations of inflammatory bowel disease (IBD, including common immune-mediated pathways, frequency, clinical course and therapy. Musculoskeletal complications are frequent and well-recognized manifestations in IBD, and affect up to 33% of patients with IBD. The strong link between the bowel and the osteo-articular system is suggested by many clinical and experimental observations, notably in HLA-B27 transgenic rats. The autoimmune pathogenic mechanisms shared by IBD and spondyloarthropathies include genetic susceptibility to abnormal antigen presentation, aberrant recognition of self, the presence of autoantibodies against specific antigens shared by the colon and other extra-colonic tissues, and increased intestinal permeability. The response against microorganisms may have an important role through molecular mimicry and other mechanisms. Rheumatic manifestations of IBD have been divided into peripheral arthritis, and axial involvement, including sacroiliitis, with or without spondylitis, similar to idiopathic ankylosing spondylitis. Other periarticular features can occur, including enthesopathy, tendonitis, clubbing, periostitis, and granulomatous lesions of joints and bones. Osteoporosis and osteomalacia secondary to IBD and iatrogenic complications can also occur. The management of the rheumatic manifestations of IBD consists of physical therapy in combination with local injection of corticosteroids and nonsteroidal anti-inflammatory drugs; caution is in order however, because of their possible harmful effects on intestinal integrity, permeability, and even on gut inflammation. Sulfasalazine, methotrexate, azathioprine, cyclosporine and leflunomide should be used for selected indications. In some cases, tumor necrosis factor-? blocking agents should be considered as first-line therapy.

Tatiana Sofía Rodríguez-Reyna, Cynthia Martínez-Reyes, Jesús Kazúo Yamamoto-Furusho

2009-11-01

 
 
 
 
401

Functional conservation between rodents and chicken of regulatory sequences driving skeletal muscle gene expression in transgenic chickens  

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Full Text Available Abstract Background Regulatory elements that control expression of specific genes during development have been shown in many cases to contain functionally-conserved modules that can be transferred between species and direct gene expression in a comparable developmental pattern. An example of such a module has been identified at the rat myosin light chain (MLC 1/3 locus, which has been well characterised in transgenic mouse studies. This locus contains two promoters encoding two alternatively spliced isoforms of alkali myosin light chain. These promoters are differentially regulated during development through the activity of two enhancer elements. The MLC3 promoter alone has been shown to confer expression of a reporter gene in skeletal and cardiac muscle in transgenic mice and the addition of the downstream MLC enhancer increased expression levels in skeletal muscle. We asked whether this regulatory module, sufficient for striated muscle gene expression in the mouse, would drive expression in similar domains in the chicken. Results We have observed that a conserved downstream MLC enhancer is present in the chicken MLC locus. We found that the rat MLC1/3 regulatory elements were transcriptionally active in chick skeletal muscle primary cultures. We observed that a single copy lentiviral insert containing this regulatory cassette was able to drive expression of a lacZ reporter gene in the fast-fibres of skeletal muscle in chicken in three independent transgenic chicken lines in a pattern similar to the endogenous MLC locus. Reporter gene expression in cardiac muscle tissues was not observed for any of these lines. Conclusions From these results we conclude that skeletal expression from this regulatory module is conserved in a genomic context between rodents and chickens. This transgenic module will be useful in future investigations of muscle development in avian species.

Taylor Lorna

2010-02-01

402

Isotype switching by a microinjected mu immunoglobulin heavy chain gene in transgenic mice.  

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Immunization of transgenic mice carrying an immunoglobulin mu heavy chain resulted in a response dominated by expression of the transgene variable region. Unexpectedly, in a large proportion of the antibody produced by immunized mice, the transgene variable region was associated with IgG rather than IgM. This demonstrates that the transgene can undergo an isotype switch. Four transgenic founder lines all exhibited transgene isotype switching despite the likelihood of random chromosomal integr...

Durdik, J.; Gerstein, R. M.; Rath, S.; Robbins, P. F.; Nisonoff, A.; Selsing, E.

1989-01-01

403

Transgenic DNA integrated into the oat genome is frequently interspersed by host DNA  

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Integration of transgenic DNA into the plant genome was investigated in 13 transgenic oat (Avena sativa L.) lines produced using microprojectile bombardment with one or two cotransformed plasmids. In all transformation events, the transgenic DNA integrated into the plant genome consisted of intact transgene copies that were accompanied by multiple, rearranged, and/or truncated transgene fragments. All fragments of transgenic DNA cosegregated, indicating that they were integrated at single gen...

Pawlowski, Wojciech P.; Somers, David A.

1998-01-01

404

Synthesis of minus-strand copies of a viral transgene during viral infections of transgenic plants  

Science.gov (United States)

Plants can be genetically engineered to express viral sequences, often resulting in resistance to the virus from which the sequence was derived. The generally accepted mechanism for this pathogen induced resistance is gene silencing. Previous work has demonstrated that viral transgenes can be incorp...

405

Characterization of a Maize Wip1 Promoter in Transgenic Plants  

Directory of Open Access Journals (Sweden)

Full Text Available The Maize Wip1 gene encodes a wound-induced Bowman-Birk inhibitor (BBI protein which is a type of serine protease inhibitor, and its expression is induced by wounding or infection, conferring resistance against pathogens and pests. In this study, the maize Wip1 promoter was isolated and its function was analyzed. Different truncated Wip1 promoters were fused upstream of the GUS reporter gene and transformed into Arabidopsis, tobacco and rice plants. We found that (1 several truncated maize Wip1 promoters led to strong GUS activities in both transgenic Arabidopsis and tobacco leaves, whereas low GUS activity was detected in transgenic rice leaves; (2 the Wip1 promoter was not wound-induced in transgenic tobacco leaves, but was induced by wounding in transgenic rice leaves; (3 the truncated Wip1 promoter had different activity in different organs of transgenic tobacco plants; (4 the transgenic plant leaves containing different truncated Wip1 promoters had low GUS transcripts, even though high GUS protein level and GUS activities were observed; (5 there was one transcription start site of Wip1 gene in maize and two transcription start sites of GUS in Wip1::GUS transgenic lines; (6 the adjacent 35S promoter which is present in the transformation vectors enhanced the activity of the truncated Wip1 promoters in transgenic tobacco leaves, but did not influence the disability of truncated Wip1231 promoter to respond to wounding signals. We speculate that an ACAAAA hexamer, several CAA trimers and several elements similar to ACAATTAC octamer in the 5'-untranslated region might contribute to the strong GUS activity in Wip1231 transgenic lines, meanwhile, compared to the 5'-untranslated region from Wip1231 transgenic lines, the additional upstream open reading frames (uORFs in the 5'-untranslated region from Wip1737 transgenic lines might contribute to the lower level of GUS transcript and GUS activity.

Shengxue Zhang

2013-12-01

406

Transgene detection by digital droplet PCR.  

Science.gov (United States)

Somatic gene therapy is a promising tool for the treatment of severe diseases. Because of its abuse potential for performance enhancement in sports, the World Anti-Doping Agency (WADA) included the term 'gene doping' in the official list of banned substances and methods in 2004. Several nested PCR or qPCR-based strategies have been proposed that aim at detecting long-term presence of transgene in blood, but these strategies are hampered by technical limitations. We developed a digital droplet PCR (ddPCR) protocol for Insulin-Like Growth Factor 1 (IGF1) detection and demonstrated its applicability monitoring 6 mice injected into skeletal muscle with AAV9-IGF1 elements and 2 controls over a 33-day period. A duplex ddPCR protocol for simultaneous detection of Insulin-Like Growth Factor 1 (IGF1) and Erythropoietin (EPO) transgenic elements was created. A new DNA extraction procedure with target-orientated usage of restriction enzymes including on-column DNA-digestion was established. In vivo data revealed that IGF1 transgenic elements could be reliably detected for a 33-day period in DNA extracted from whole blood. In vitro data indicated feasibility of IGF1 and EPO detection by duplex ddPCR with high reliability and sensitivity. On-column DNA-digestion allowed for significantly improved target detection in downstream PCR-based approaches. As ddPCR provides absolute quantification, it ensures excellent day-to-day reproducibility. Therefore, we expect this technique to be used in diagnosing and monitoring of viral and bacterial infection, in detecting mutated DNA sequences as well as profiling for the presence of foreign genetic material in elite athletes in the future. PMID:25375130

Moser, Dirk A; Braga, Luca; Raso, Andrea; Zacchigna, Serena; Giacca, Mauro; Simon, Perikles

2014-01-01

407

Human prion strain selection in transgenic mice  

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Transgenic (Tg) mice expressing chimeras of mouse and human prion proteins (PrP) have shorter incubation periods for Creutzfeldt-Jakob disease (CJD) prions than mice expressing full-length human PrP. Increasing the sequence similarity of the chimeric PrP to mouse PrP, by reverting human residues to mouse, resulted in a Tg line, denoted Tg22372, which was susceptible to sporadic (s) CJD prions in ~110 days 1. Reversion of one additional residue (M111V) resulted in a new Tg line, termed Tg1014,...

Giles, Kurt; Glidden, David V.; Patel, Smita; Korth, Carsten; Groth, Darlene; Lemus, Azucena; Dearmond, Stephen J.; Prusiner, Stanley B.

2010-01-01

408

A transgenic tri-modality reporter mouse.  

Science.gov (United States)

Transgenic mouse with a stably integrated reporter gene(s) can be a valuable resource for obtaining uniformly labeled stem cells, tissues, and organs for various applications. We have generated a transgenic mouse model that ubiquitously expresses a tri-fusion reporter gene (fluc2-tdTomato-ttk) driven by a constitutive chicken ?-actin promoter. This "Tri-Modality Reporter Mouse" system allows one to isolate most cells from this donor mouse and image them for bioluminescent (fluc2), fluorescent (tdTomato), and positron emission tomography (PET) (ttk) modalities. Transgenic colonies with different levels of tri-fusion reporter gene expression showed a linear correlation between all three-reporter proteins (R(2)=0.89 for TdTomato vs Fluc, R(2)=0.94 for Fluc vs TTK, R(2)=0.89 for TdTomato vs TTK) in vitro from tissue lysates and in vivo by optical and PET imaging. Mesenchymal stem cells (MSCs) isolated from this transgenics showed high level of reporter gene expression, which linearly correlated with the cell numbers (R(2)=0.99 for bioluminescence imaging (BLI)). Both BLI (R(2)=0.93) and micro-PET (R(2)=0.94) imaging of the subcutaneous implants of Tri-Modality Reporter Mouse derived MSCs in nude mice showed linear correlation with the cell numbers and across different imaging modalities (R(2)=0.97). Serial imaging of MSCs transplanted to mice with acute myocardial infarction (MI) by intramyocardial injection exhibited significantly higher signals in MI heart at days 2, 3, 4, and 7 (p<0.01). MSCs transplanted to the ischemic hindlimb of nude mice showed significantly higher BLI and PET signals in the first 2 weeks that dropped by 4(th) week due to poor cell survival. However, laser Doppler perfusion imaging revealed that blood circulation in the ischemic limb was significantly improved in the MSCs transplantation group compared with the control group. In summary, this mouse can be used as a source of donor cells and organs in various research areas such as stem cell research, tissue engineering research, and organ transplantation. PMID:23951359

Yan, Xinrui; Ray, Pritha; Paulmurugan, Ramasamy; Tong, Ricky; Gong, Yongquan; Sathirachinda, Ataya; Wu, Joseph C; Gambhir, Sanjiv S

2013-01-01

409

Sequence similarity between the cp gene and the transgene in transgenic papayas = Similaridade de seqüência entre o gene cp do vírus e do transgene presente em mamoeiros transgênicos  

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The Papaya ringspot virus (PRSV) coat protein transgene present in 'Rainbow' and 'SunUp' papayas disclose high sequence similarity (>89%) to the cp gene from PRSV BR and TH. Despite this, both isolates are able to break down the resistance in 'Rainbow', while only the latter is able to do so in 'SunUp'. The objective of this work was to evaluate the degree of sequence similarity between the cp gene in the challenge isolate and the cp transgene in transgenic papayas resistant to PRSV. The prod...

Souza, M. T.; Teixeira, M.; Gonsalves, D.

2005-01-01

410

Effect of stacking insecticidal cry and herbicide tolerance epsps transgenes on transgenic maize proteome.  

Science.gov (United States)

BackgroundThe safe use of stacked transgenic crops in agriculture requires their environmental and health risk assessment, through which unintended adverse effects are examined prior to their release in the environment. Molecular profiling techniques can be considered useful tools to address emerging biosafety gaps. Here we report the first results of a proteomic profiling coupled to transgene transcript expression analysis of a stacked commercial maize hybrid containing insecticidal and herbicide tolerant traits in comparison to the single event hybrids in the same genetic background.ResultsOur results show that stacked genetically modified (GM) genotypes were clustered together and distant from other genotypes analyzed by PCA. Twenty-two proteins were shown to be differentially modulated in stacked and single GM events versus non-GM isogenic maize and a landrace variety with Brazilian genetic background. Enrichment analysis of these proteins provided insight into two major metabolic pathway alterations: energy/carbohydrate and detoxification metabolism. Furthermore, stacked transgene transcript levels had a significant reduction of about 34% when compared to single event hybrid varieties.ConclusionsStacking two transgenic inserts into the genome of one GM maize hybrid variety may impact the overall expression of endogenous genes. Observed protein changes differ significantly from those of single event lines and a conventional counterpart. Some of the protein modulation did not fall within the range of the natural variability for the landrace used in this study. Higher expression levels of proteins related to the energy/carbohydrate metabolism suggest that the energetic homeostasis in stacked versus single event hybrid varieties also differ. Upcoming global databases on outputs from ¿omics¿ analyses could provide a highly desirable benchmark for the safety assessment of stacked transgenic crop events. Accordingly, further studies should be conducted in order to address the biological relevance and implications of such changes. PMID:25490888

Agapito-Tenfen, Sarah; Vilperte, Vinicius; Benevenuto, Rafael; Rover, Carina; Traavik, Terje; Nodari, Rubens

2014-12-10

411

Microarray analysis of androgen-regulated gene expression in testis: the use of the androgen-binding protein (ABP)-transgenic mouse as a model  

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Abstract Background Spermatogenesis is an androgen-dependent process, yet the molecular mechanisms of androgens' actions in testis are poorly understood. Transgenic mice overexpressing rat androgen-binding protein (ABP) in their testes have reduced levels of intratesticular androgens and, as a result, show a progressive impairment of spermatogenesis. We used this model to characterize changes in global gene expression in testis in response to reduced bioavailability of androg...

Grossman Gail; Jeyaraj Durairaj A; Petrusz Peter

2005-01-01

412

Calcitonin gene-related peptide-evoked sustained tachycardia in calcitonin receptor-like receptor transgenic mice is mediated by sympathetic activity  

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Calcitonin gene-related peptide (CGRP) and adrenomedullin (AM) are potent vasodilators and exert positive chronotropic and inotropic effects on the heart. Receptors for CGRP and AM are calcitonin receptor-like receptor (CLR)/receptor-activity-modifying protein (RAMP) 1 and CLR/RAMP2 heterodimers, respectively. The present study was designed to delineate distinct cardiovascular effects of CGRP and AM. Thus a V5-tagged rat CLR was expressed in transgenic mice in the vascular musculature, a reco...

Kunz, T. H.; Scott, M.; Ittner, L. M.; Fischer, J. A.; Born, W.; Vogel, J.

2007-01-01

413

Identification of brain-derived neurotrophic factor promoter regions mediating tissue-specific, axotomy-, and neuronal activity-induced expression in transgenic mice  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The structure of rat brain-derived neurotrophic factor (BDNF) gene is complex; four 5' exons are linked to separate promoters and one 3' exon is encoding the BDNF protein. To analyze the relative importance of the regulatory regions in vivo, we have generated transgenic mice with six different promoter constructs of the BDNF gene fused to the chloramphenicol acetyl transferase reporter gene. High level and neuronal expression of the reporter gene, that in many respects recapitulated BDNF gene...

1995-01-01

414

Pituitary-directed leukemia inhibitory factor transgene forms Rathke's cleft cysts and impairs adult pituitary function. A model for human pituitary Rathke's cysts.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Leukemia inhibitory factor (LIF) and LIF receptors are expressed in adenohypophyseal cells and LIF regulates pituitary hormone transcription and cell replication in vitro. Therefore, transgenic mice expressing pituitary-directed LIF driven by the rat growth hormone (GH) promoter were generated to evaluate the impact of LIF on pituitary development. Three founders were established with diminished linear growth and body weight (57-65% of wild type [WT]), and intense anterior pituitary LIF immun...

Akita, S.; Readhead, C.; Stefaneanu, L.; Fine, J.; Tampanaru-sarmesiu, A.; Kovacs, K.; Melmed, S.

1997-01-01

415

Overview of the investigation of transgenic plums in Romania  

Science.gov (United States)

Transgenic plums of Prunus domestica L. transformed with the Plum pox virus coat protein gene (PPV-CP) were the subjects of three experiments undertaken in Romania. In the first experiment, PPV-CP transgenic