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1

Lactobacillus GG prevents recurrence of colitis in HLA-B27 transgenic rats after antibiotic treatment  

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Background and aims: Bacteroides vulgatus induces colitis in gnotobiotic HLA-B27 transgenic (TG) rats while broad spectrum antibiotics prevent and treat colitis in specific pathogen free (SPF) TG rats although disease recurs after treatment ends. Lactobacilli treat human pouchitis and experimental colitis. We investigated if Lactobacillus rhamnosus GG (L GG) can prevent colitis in TG rats monoassociated with B vulgatus and if L GG or Lactobacillus plantarum 299v (LP 299v) can treat establishe...

Dieleman, L. A.; Goerres, M. S.; Arends, A.; Sprengers, D.; Torrice, C.; Hoentjen, F.; Grenther, W. B.; Sartor, R. B.

2003-01-01

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Transfer of the inflammatory disease of HLA-B27 transgenic rats by bone marrow engraftment  

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We have previously produced lines of rats transgenic for HLA-B27 and human beta 2-microglobulin (h beta 2m) that develop a progressive inflammatory disease sharing many clinical and histologic features with the B27-associated human spondyloarthropathies, including gut and male genital inflammation, arthritis, and psoriasiform skin lesions. Other transgenic lines that express lower levels of B27 and h beta 2m remain healthy. To investigate the cellular basis for the multisystem inflammatory di...

1993-01-01

3

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

4

Autoimmune Epididymo-orchitis is Essential to the Pathogenesis of Male-Specific Spondyloarthritis in HLA-B27 Transgenic Rats  

Science.gov (United States)

Objective Male rats transgenic for HLA-B27 and human-?2-microglobulin (Hu-?2m) spontaneously develop epididymo-orchitis preceding spondyloarthritis. In the specific B27/Hu-?2m transgenic cross (21-3x382-2)F1, only the males develop spondyloarthritis, and neither sex develops gut inflammation. We asked whether epididymo-orchitis and spondyloarthritis in the (21-3x382-2)F1 males are causally related, and we characterized the epididymo-orchitis. Methods B27/Hu-?2m (21-3x382-2)F1 transgenic males underwent bilateral, unilateral, or sham epididymo-orchiectomy between ages 36 and 125 d. Castrated rats were given testosterone replacement. Alternatively, the 21-3 and 283-2 transgene loci were crossed with a transgene inducing aspermatogenesis. Rats were observed for epididymo-orchitis, arthritis, and spondylitis. Results In unmanipulated transgenic rats, inflammation was first evident in the ductuli efferentes (DE, ducts linking rete testis to epididymis) as early as age 30 d. The inflammation was initially neutrophilic, and later became granulomatous. Serum anti-sperm and anti-testis cell antibodies appeared after age 70 d. Cells infiltrating the testes were predominantly CD4+ T cells and CD68+ or CD163+ macrophages. Quantitative PCR of DE, epididymis, and testis showed elevations of IFN?, IL-10, andIL-17A. IL-12A, IL-22, IL-23A, and IL-23R were also examined in DE and found elevated. Remarkably, castration before 91 d of age completely prevented subsequent arthritis and spondylitis, as did transgene-induced azospermia. Conclusion In the (21-3x283-2)F1 HLA-B27/Hu-?2m transgenic rats, autoimmune epididymo-orchitis develops spontaneously at 30 d, the age when antigen-positive meiotic germ cells first exit the testis. Persistent testicular inflammation and/or antigenic stimulation are essential prerequisites to subsequent spondyloarthritis. Dysregulated innate immunity at immune privileged sites may be an essential mechanism triggering spondyloarthritis.

Taurog, Joel D.; Rival, Claudia; van Duivenvoorde, Leonie M.; Satumtira, Nimman; Dorris, Martha L.; Sun, Margaret; Shelton, John M.; Richardson, James A.; Hamra, F. Kent; Hammer, Robert E.; Tung, Kenneth S. K.

2012-01-01

5

HLA-B27 misfolding and ankylosing spondylitis.  

Science.gov (United States)

Understanding how HLA-B27 contributes to the pathogenesis of spondyloarthritis continues to be an important goal. Current efforts are aimed largely on three areas of investigation; peptide presentation to CD8T cells, abnormal forms of the HLA-B27 heavy chain and their recognition by leukocyte immunoglobulin-like receptors on immune effector cells, and HLA-B27 heavy chain misfolding and intrinsic biological effects on affected cells. In this chapter we review our current understanding of the causes and consequences of HLA-B27 misfolding, which can be defined biochemically as a propensity to oligomerize and form complexes in the endoplasmic reticulum (ER) with the chaperone BiP (HSPA5/GRP78). HLA-B27 misfolding is linked to an unusual combination of polymorphisms that identify this allele, and cause the heavy chain to fold and load peptides inefficiently. Misfolding can result in ER-associated degradation (ERAD) of heavy chains, which is mediated in part by the E3 ubiquitin ligase HRD1 (SYVN1), and the ubiquitin conjugating enzyme UBE2JL. Upregulation of HLA-B27 and accumulation of misfolded heavy chains can activate ER stress signaling pathways that orchestrate the unfolded protein response. In transgenic rats where HLA-B27 is overexpressed, UPR activation is prominent. However, it is specific for heavy chain misfolding, since overexpression of HLA-B7, an allele that does not misfold, fails to generate ER stress. UPR activation has been linked to cytokine dysregulation, promoting lL-23, IFN?, and lL-1? production, and may activate the IL-23/IL-17 axis in these rats. IL-1? and IFN? are pro- and anti-osteoclastogenic cytokines, respectively, that modulate osteoclast development in HLA-B27-expressing transgenic rat monocytes. Translational studies of patient derived cells expressing HLA-B27 at physiologic levels have provided evidence that ER stress and UPR activation can occur in peripheral blood, but this has not been reported to date in isolated macrophages. Inflamed gastrointestinal tissue reveals evidence for HLA-B27 misfolding, ERAD, and autophagy, without acute UPR activation. A more complete picture of conditions that impact HLA-B27 folding and misfolding, the full spectrum and time course of consequences of ER stress, and critical cell types involved is needed to understand the role of HLA-B27 misfolding in spondyloarthritis pathogenesis. PMID:23993278

Colbert, Robert A; Tran, Tri M; Layh-Schmitt, Gerlinde

2014-01-01

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Rat MHC-linked peptide transporter alleles strongly influence peptide binding by HLA-B27 but not B27-associated inflammatory disease  

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Rats transgenic for the human MHC molecule HLA-B27 were used to study the effect of two alleles, cim{sup a} and cim{sup b}, which are associated with peptide transport by the MHC-encoded Tap2 transporter, on the function of HLA-B27 as a restriction element for CTL recognition of the male H-Y minor H Ag and on the multisystem inflammatory disease characteristic of B27 transgenic rats. Anti-H-Y CTL generated in cim{sup a} B27 transgenic rats lysed male B27 cim{sup b/b} targets significantly less well than cim{sup a/a} or cim{sup a/b} targets. Addition of exogenous H-Y peptides to male B27 cim{sup b/b} targets increased susceptibility to lysis to the level of cim{sup a/a} targets sensitized with exogenous H-Y peptides. {sup 3}H-labeled peptides eluted from B27 molecules of lymphoblasts from rats of two cim{sup b} and three cim{sup a} RT1 haplotypes showed that the cim{sup b} peptide pool favors comparatively longer and/or more hydrophobic peptides. These results indicate that RT1-linked Tap2 polymorphism in the rat strongly influences peptide loading of HLA-B27. Nonetheless, the prevalence and severity of multisystem inflammatory lesions were comparable in backcross rats bearing either cim{sup a/b} or cim{sup b/b}. It thus appears either that binding of specific peptides to B27 is unimportant in the pathogenesis of B27-associated disease or that the critical peptides, unlike H-Y and many others, are not influenced by Tap transporter polymorphism. 42 refs., 6 figs., 3 tabs.

Simmons, W.A.; Satumtira, Nimman; Taurog, J.D. [Univ. of Texas Southwestern Medical Center, Dallas, TX (United States)] [and others

1996-02-15

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Seronegative pauciarticular arthritis and HLA B27.  

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Twenty-six patients with a pauciarticular arthritis have been studied clinically, radiologically and with histocompatibility typing. An increased frequency of HLA B27 was found (p = 1.87 x 10(-12)). Low back and buttock pain, Achilles tendinitis and dactylitis of the toes were more frequent in HLA-B27 positive patients. It is suggested that histocompatibility testing may be of some value in diagnosis and in the investigation of the possible 'reactive' nature of this type of arthritis.

Eastmond, C. J.; Rajah, S. M.; Tovey, D.; Wright, V.

1980-01-01

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HLA B27 y las espondilartropatías seronegativas  

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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.

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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

1995-01-01

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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 pat...

Huhtinen, M.; Karma, A.

2000-01-01

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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 sero...

Kerkhoff, F.; Rothova, A.

2000-01-01

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

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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.

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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;...

Huppertz, H.; Heesemann, J.

1997-01-01

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Specificity of anti-HLA-B27 cytotoxic T lymphocytes.  

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Sub-types of HLA-B27 were detected by cytotoxic T lymphocytes (CTL) generated between HLA-A, -B- and -C-identical B27-positive individuals. We now report the specificity of six independent CTL's generated by mixed lymphocyte culture (MLC) of HLA-A, -B and -C serologically identical B27-positive responder and stimulator cells. Three CTL's recognize one sub-type, and three the other. The combined reactivity of all CTL's allows unequivocal "typing" of B27-positive cells for the two different sub-types B27K and B27W. The specificity of two CTL's was analysed by cold-target inhibition. The results indicate that (1) no further sub-types of HLA-B27 can be detected by the CTL's raised in these combinations; (2) the majority of the CTL's is directed against the B27 antigens; and (3) "extra reactions" on B27-negative cells are caused by a subset(s) of CTL's recognizing unknown antigens shared between stimulator and target cells. CTL's raised by stimulation of HLA-B27-negative responder cells with B27-positive cells of either sub-type lysed all B27-positive target cells indiscriminately. In cold-target inhibition, however, B27-positive cells, carrying the sub-type of B27 different from that of the stimulator, could not inhibit the lysis of cells bearing the stimulator sub-type of B27. This indicates the activation, in B27-negative responders, of at least two different groups of CTL clones, one directed against shared determinants of HLA-B27, and one against the HLA-B27 sub-type. Heterogeneity of the HLA-B27 antigen may have implications for studies on the well-known association between this antigen and various diseases. PMID:6606238

Breuning, M H; Breur, B S; Engelsma, M Y; Huis, B; Iványi, P

1983-10-01

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HLA-B27, but Not HLA-B7, Immunodominance to Influenza Is ERAP Dependent.  

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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

20

Short peptide sequence identity between human viruses and HLA-B27-binding human 'self' peptides.  

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Molecular mimicry and arthritogenic peptides form the basis of hypotheses that attempt to explain the pathogenesis of HLA-B27-positive ankylosing spondylitis (AS). We propose, therefore, that certain human viruses may possess peptide sequences that mimic HLA-B27-binding human 'self' peptides which might induce or play a significant role in AS. In the present study, we performed bioinformatic analysis, using BLASTP, of the human virus proteome and HLA-B27-binding human 'self' peptides including peptides derived from arthritogenic sequences. We identified that some HLA-B27-binding peptides, particularly those present in proteins of the cartilage and bone, are highly similar to those present in viruses known to cause chronic infection. We suggest that the identical short amino acid sequences shared between human viruses and HLA-B27 peptides may play a role in the pathogenesis of AS. PMID:24362932

Sun, Shipeng; Wang, Tao; Pang, Bo; Wei, Huamin; Liu, Guijian

2014-06-01

 
 
 
 
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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

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

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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 r...

2012-01-01

23

The clinical characteristics of retinal vasculitis in HLA-B27-positive patients.  

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Abstract Purpose: To investigate the ocular and systemic manifestations of retinal vasculitis in HLA-B27-positive patients. Methods: Retrospective noncomparative case series of 9 HLA-B27-positive patients with uveitis and retinal vasculitis. Main outcome measures consisted of ocular and angiographic findings and assessment of any additional systemic disorders. Results: Three male and 6 female HLA-B27-positive patients with a median age of 32 years were diagnosed with retinal vasculitis. Concurrent intraocular inflammation was noted in all patients. All patients suffered from extensive vasculitis of the large retinal veins. Five patients developed retinal vasculitis at the onset of uveitis and the remaining 4 exhibited retinal vasculitis 1-15 years after the onset of uveitis. Vascular occlusions occurred in 4 patients and subsequent neovascularizations developed in 3. Three patients were diagnosed with an HLA-B27-associated systemic disease. Conclusion: Retinal vasculitis may develop in the wake of HLA-B27-associated uveitis and might represent a rare manifestation of HLA-B27-associated disease. PMID:24102118

Braakenburg, Arthur Menno; Rothova, Aniki

2014-06-01

24

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 identifying at risk family members. (author)

2007-01-01

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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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HLA-B27 Selects for Rare Escape Mutations that Significantly Impair Hepatitis C Virus Replication and Require Compensatory Mutations  

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HLA-B27 is associated with spontaneous viral clearance in hepatitis C virus (HCV) infection. Viral escape within the immunodominant HLA-B27 restricted HCV-specific CD8+ T cell epitope NS5B2841-2849 (ARMILMTHF) has been shown to be limited by viral fitness costs as well as broad T cell cross-recognition, suggesting a potential mechanism of protection by HLA-B27. Here, we studied the subdominant HLA-B27 restricted epitope NS5B2936-2944 (GRAAICGKY) in order to further define the mechanisms of pr...

2011-01-01

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Punctate palmoplantar keratoderma associated with morbus Bechterew and HLA B 27. A family study.  

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Four patients in a family with punctate palmoplantar keratoderma (Buschke-Fischer) associated with Morbus Bechterew and HLA B 27 in 3 of the family members are reported. Without severe side effect, the proband was successfully treated with 50 mg etretinate per day for 6 weeks. PMID:2459882

Gamborg Nielsen, P

1988-01-01

28

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

29

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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HLA-B27 Expression Does Not Modulate Intracellular Chlamydia trachomatis Infection of Cell Lines  

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Chlamydia trachomatis is an obligate intracellular pathogen. Infection of susceptible individuals with this bacterium can trigger the development of reactive arthritis, an acute inflammation that is associated with the expression of the class I major histocompatibility antigen, HLA-B27. Other facultative intracellular pathogens, such as Yersinia and Salmonella spp., are also known triggers of reactive arthritis. Previous studies report conflicting results concerning whether the presence of HL...

Young, J. L.; Smith, L.; Matyszak, M. K.; Gaston, J. S. H.

2001-01-01

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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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Antinuclear antibody and HLA-B27 positive uveitis: combination of two diseases?  

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AIMS/BACKGROUND—Anterior uveitis associated with juvenile chronic arthritis concerns two different clinical entities: firstly, antinuclear antibody (ANA) positive patients who have a chronic anterior uveitis with severe complications and often a poor visual prognosis; secondly, usually HLA-B27 positive children, predominantly boys, with unilateral recurrent anterior uveitis. Three patients are described who had a combination of clinical and laboratory features of both diseases.?METHOD...

Bosch-driessen, E.; Lardy, N.; Rothova, A.

1997-01-01

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Association of HLA-B27 and ERAP1 with ankylosing spondylitis susceptibility in Beijing Han Chinese.  

Science.gov (United States)

This study investigated the genetic polymorphisms of HLA-B27, together with polymorphisms on endoplasmic reticulum aminopeptidase 1 (ERAP1), and susceptibility for ankylosing spondylitis (AS) in the Beijing Han population. A case-control study was carried out for 602 AS patient samples and 619 matched controls of Han Chinese. HLA-B27 genotyping was performed by polymerase chain reaction-sequence specific primers (PCR-SSP), and four ERAP1 SNPs (rs27037, rs27980, rs27582, and rs27434) were selected and genotyped on the Sequenom iPlex platform (Sequenom, San Diego, CA). Association analysis was performed using the likelihood ratio ?(2) test. This study identified four HLA-B27 alleles in Beijing Han AS patients, B*27:02, B*27:04, B*27:05, and B*27:07, of which B*27:05 was the most significant geographical different subtype among AS patients in Chinese. Our results confirmed that HLA-B27 was strongly associated with AS (P=1.9 × 10(-150) ), and the most strongly associated alleles were B*27:04, B*27:05, and B*27:02. Our study also confirmed a weak association between ERAP1 (rs27434) and AS. We also observed that for HLA-B*27:02 and HLA-B*27:04 positive AS patients, rs27434 and rs27582 were associated with AS. In contrast, for HLA-B27-negative and HLA-B*27:05-positive AS patients, this association was not observed. This is the first study to show that both B27 and ERAP1 are AS genetic susceptibility genes in Beijing Han. Interactions between ERAP1 and HLA-B*27:02 and B*27:04 may play an important role in the AS pathogenesis. PMID:24666027

Zhang, Z; Dai, D; Yu, K; Yuan, F; Jin, J; Ding, L; Hao, Y; Liang, F; Liu, N; Zhao, X; Long, J; Xi, Y; Sun, Y-Y

2014-05-01

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Functional variants of ERAP1 gene are associated with HLA-B27 positive spondyloarthritis.  

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We investigated two nonsynonymous variants (rs30187 and rs27044) of ERAP1 gene in HLA-B27 positive individuals (150 spondyloarthritis and 108 controls) and in general ankylosing spondylitis (AS) patients (n = 137) vs random controls (n = 139). Both single nucleotide polymorphisms (SNPs) were associated with the risk of spondyloarthritis [odds ratio (OR) 1.80, 95% confidence interval (CI) 1.24-2.62, P = 0.001 for rs30187, OR 1.58, 95% CI 1.07-2.34, P = 0.02 for rs27044]. The CC haplotype was a protective factor (P = 0.002), while the TG haplotype was a risk factor (P = 0.01) for spondyloarthritis. The SNP rs30187 was also associated with the risk of HLA-B27+ AS. For the general group of AS, the carriers of minor alleles showed an increased risk for the disease (OR 1.92, 95% CI 1.17-3.13 for rs30187, OR 1.74, 95% CI 1.08-2.80 for rs27044). This is the first study that shows the association of ERAP1 gene variants and haplotypes with HLA-B27 positive spondyloarthritis. PMID:23800305

Cherciu, M; Popa, L O; Bojinca, M; Dutescu, M I; Bojinca, V; Bara, C; Popa, O M

2013-09-01

35

Subtypes of HLA-B27 detected by cytotoxic T lymphocytes and their role in self-recognition.  

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In the present study cytotoxic T lymphocytes were generated in MLC of lymphocytes from two unrelated HLA-A, B, C-identical, B27-positive, but D/DR-different, individuals. These CTL were shown to detect subtypes of HLA-B27. CTL specific for influenza virus lysed infected target cells matched for HLA-B27 only when they shared the same subtype. This indicates that the two subtypes of HLA-B27 detected by CTL function also as distinct elements in a self-restricted CTL response. Both subtypes were found among patients with ankylosing spondylitis. PMID:6186653

Breuning, M H; Lucas, C J; Breur, B S; Engelsma, M Y; de Lange, G G; Dekker, A J; Biddison, W E; Ivanyi, P

1982-12-01

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Ultrasonographic Assessment of Enthesitis in HLA-B27 Positive Patients with Rheumatoid Arthritis, a Matched Case-Only Study  

Science.gov (United States)

Introduction HLA-B27 has a modifier effect on the phenotype of multiple diseases, both associated and non-associated with it. Among these effects, an increased frequency of clinical enthesitis in patients with Rheumatoid Arthritis (RA) has been reported but never explored again. We aimed to replicate this study with a sensitive and quantitative assessment of enthesitis by using standardized ultrasonography (US). Methods The Madrid Sonography Enthesitis Index (MASEI) was applied to the US assessment of 41 HLA-B27 positive and 41 matched HLA-B27 negative patients with longstanding RA. Clinical characteristics including explorations aimed to evaluate spondyloarthrtitis and laboratory tests were also done. Results A significant degree of abnormalities in the entheses of the patients with RA were found, but the MASEI values, and each of its components including the Doppler signal, were similar in HLA-B27 positive and negative patients. An increase of the MASEI scores with age was identified. Differences in two clinical features were found: a lower prevalence of rheumatoid factor and a more common story of low back pain in the HLA-B27 positive patients than in the negative. The latter was accompanied by radiographic sacroiliitis in two HLA-B27 positive patients. No other differences were detected. Conclusion We have found that HLA-B27 positive patients with RA do not have more enthesitis as assessed with US than the patients lacking this HLA allele. However, HLA-B27 could be shaping the RA phenotype towards RF seronegativity and axial involvement.

Mera-Varela, Antonio; Ferreiro-Iglesias, Aida; Perez-Pampin, Eva; Porto-Silva, Marisol; Gomez-Reino, Juan J.; Gonzalez, Antonio

2013-01-01

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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

1983-01-01

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

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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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CD8+ T-cell autoreactivity to an HLA-B27–restricted self-epitope correlates with ankylosing spondylitis  

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HLA-B27 is highly associated with ankylosing spondylitis (AS), but the mechanism is unknown. Among the HLA-B27 alleles, B*2709, which differs by one amino acid from the susceptible B*2705, is not associated with the disease. Here, we analyze the reactivity, in patients with AS and in healthy controls carrying the B*2709 or B*2705 alleles, to an EBV epitope derived from LMP2 (236-244) and to a sequence-related self-peptide from vasoactive intestinal peptide receptor 1 (VIP1R 400-408). We found...

Fiorillo, Maria T.; Maragno, Monica; Butler, Richard; Dupuis, Maria L.; Sorrentino, Rosa

2000-01-01

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Low T cell production of TNF? and IFN? in ankylosing spondylitis: its relation to HLA-B27 and influence of the TNF-308 gene polymorphism  

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OBJECTIVE—To test the hypothesis that ankylosing spondylitis (AS) is a T helper cell type 2 polarised disease by quantifying the T cell cytokines interferon ? (IFN?), interleukin 4 (IL4), tumour necrosis factor ? (TNF?), and IL10 at the single cell level in patients with AS in comparison with healthy HLA-B27 negative and HLA-B27 positive controls.?METHODS—Peripheral blood mononuclear cells from 65 subjects (25 HLA-B27 positive patients with active AS, 18 healthy HLA-B27 p...

2001-01-01

 
 
 
 
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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.

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

42

HLA B27 and defects in the T-cell system in Whipple's disease.  

Science.gov (United States)

The cellular immune system was tested in nine patients with Whipples' disease. Three patients had active disease, and six had been in remission for up to 10 years. Intradermal delayed hypersensitivity reactions to candidin, trichophytin, tuberculin and varidase, T-cell counts as determined by E-rosettes, allogeneic stimulation of lymphocytes in the mixed lymphocyte culture, and mitogenic activation of lymphocytes by concanavalin A, phytohaemagglutinin and by pokeweed mitogen, were tested in the patients and compared with control subjects. HLA typing was performed in all patients. The reaction to tuberculin and varidase, the T-cell counts and the activation of lymphocytes by concanavalin A were significantly reduced in patients with active disease and in patients during remission. The reaction to candidin and trichophytin was poor even in the controls. The mean results of the mixed lymphocyte culture, phytohaemagglutinin, and pokeweed mitogen activation tests were not significantly different from the controls. In patients with active disease the mixed lymphocyte culture reaction and the T-cell counts were less than in patients in remission. The results suggest a persistent defect of T-cells in patients with Whipple's disease, a defect that is more severe in patients with active disease. The finding of HLA B27 in four of thenine patients supports the hypothesis of primary rather than secondary impairment of the cellular immune system in Whipple's disease. PMID:93049

Feurle, G E; Dörken, B; Schöpf, E; Lenhard, V

1979-10-01

43

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

1987-11-15

44

Expression of arthritis-causing HLA-B27 on Hela cells promotes induction of c-fos in response to in vitro invasion by Salmonella typhimurium.  

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HLA-B27 confers a very strong genetic predisposition to development of a reactive arthritis after infection by bacteria such as Salmonella typhimurium. This study examines the role of HLA-B27 in the initiation of the earliest host activities after exposure to Salmonella, namely activation of the immediate early genes in the epithelial cells. Our major finding is that in Hela cells, the expression of c-fos was induced by Salmonella invasion only when the cells expressed the transfected HLA-B27...

Ikawa, T.; Ikeda, M.; Yamaguchi, A.; Tsai, W. C.; Tamura, N.; Seta, N.; Trucksess, M.; Raybourne, R. B.; Yu, D. T.

1998-01-01

45

HLA-B27-Transfected (Salmonella Permissive) and HLA-A2-Transfected (Salmonella Nonpermissive) Human Monocytic U937 Cells Differ in Their Production of Cytokines  

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The cytokine secretion of the Salmonella-permissive, HLA-B27-positive U937 cells was examined, as it was previously shown that these cells kill Salmonella less efficiently than controls. Salmonella-permissive U937 cells showed upregulated production of interleukin 10 and to a lesser extent tumor necrosis factor alpha. HLA-B27-associated modulation of cytokine responses may have importance in the pathogenesis of reactive arthritis.

Ekman, Pa?ivi; Saarinen, Marja; He, Qiushui; Gripenberg-lerche, Christel; Gro?nberg, Alvar; Arvilommi, Heikki; Granfors, Kaisa

2002-01-01

46

Effect of HLA-B*27 and its subtypes on clinical manifestations and severity of ankylosing spondylitis in Iranian patients.  

Science.gov (United States)

The aim of this study was to assess the role of HLA-B*27 and it's subtypes in determining severity and clinical manifestations of ankylosing spondylitis (AS).A total of 163 AS patients were assessed for clinical manifestations and severity using structured questionnaires. HLA-B*27 screening and B*27 sub-typing were performed by PCR.One hundred twenty two patients (74.8%) were B*27 positive. The male to female ratio, peripheral arthritis, steroid use, intense dorsal kyphosis and decrease of cervical slope had a significantly higher frequency in B*27 positive patients compared to B*27 negative ones (p=0.01, 0.001, 0.01, 0.04 and 0.04, respectively). However, the age of diagnosis was significantly lower in B*27 positive patients (p=0.005). Trend in uveitis and some severity markers including: BASMI and ASQoL were toward higher values in B*27 positive group with no significant difference. After controlling confounding variables, significant relationship was found only between B*27 and BASMI (p=0.01). B*27 subtypes in patients were included B*2705: 48.4%, B*2702: 42.6%, B*2704: 5.7% and B*2707: 3.3%. No significant differences were seen for severity markers and clinical manifestations between subtypes; although trend toward lower values of severity markers, less intense dorsal kyphosis and less decrease of cervical slope were observed in B*2704 and B*2707 versus other polymorphisms.Clinical features and severity of AS is influenced by HLA-B*27. Trend toward higher severity markers in B*2705 and B*2702 versus other polymorphisms might be subject of interest for evaluation in other ethnicities with concentration to other novel susceptibility genes co-inherited in each B*27 subtype. PMID:23996708

Fallahi, Sasan; Mahmoudi, Mahdi; Nicknam, Mohammad Hossein; Gharibdoost, Farhad; Farhadi, Elham; Saei, Azad; Nourijelyani, Keramat; Ahmadzadeh, Nooshin; Jamshidi, Ahmad Reza

2013-12-01

47

HLA-B27 polymorphism in patients with juvenile and adult-onset ankylosing spondylitis in Southern China.  

Science.gov (United States)

Distribution of B27 subtypes in juvenile and adult-onset ankylosing spondylitis (JAS and AAS) in Southern China was studied. A total of 505 patients belonged to Han population were included (145 JAS and 360 AAS patients), and 1368 healthy individuals were included as controls. Human leukocyte antigen (HLA)-B27 typing was performed by Luminex liquid array combining polymerase chain reaction-sequence specific oligonucleotide probe (PCR-SSOP) and/or serological method. HLA-B27 subtyping was performed by polymerase chain reaction-sequence specific primer (PCR-SSP). The sequence-based typing was performed for the B*2715 samples to verify the PCR-SSP results. HLA-B27 was presented in 453 of 505 patients (89.7%), compared with 74 of 1368 controls (5.41%). B*2704 subtype in AS group was significantly higher than controls and B*2705 subtype significantly lower. B*2715 and B*2702 were found in 1.32% and 0.66% of the B27-positive patients but none in controls, and there was no significant difference between either of them and controls. B27-positive patients were 134 (92.4%) in JAS group and 319 (88.6%) in AAS group. There was no significant difference for B27 subtypes distribution between JAS (B*2704, 05, 15) and AAS (B*2704, 05, 15, 02) groups. The frequency of B*2715 in two groups was 3 (2.24%) and 3 (0.94%), respectively. The onset age of three JAS patients carrying B*2715 was 5, 9 and 13 years old, respectively. Our results suggested that B*2704 was the predominant subtype in AS patients in Southern China. B*2715 was observed in AS group only and slightly more in JAS than in AAS, and the patients carrying this allele tended to have early onset, B*2715 may be disease-association subtype. PMID:20196819

Mou, Y; Wu, Z; Gu, J; Liao, Z; Lin, Z; Wei, Q; Huang, J; Li, Q

2010-01-01

48

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

49

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

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[Clinical, radiographic and biologic particularities of ankylosing spondylitis in Tunisian patients according to the presence or the absence of the HLA B27 and its sub-types].  

Science.gov (United States)

To assess the clinical, radiographic and biologic particularities of ankylosing spondylitis (AS) in Tunisian patients according to HLA B27 and its sub-types statute. This was a case-control study that included 100 patients (85 males/15 females) with AS according to the modified New York criteria. Demographic, clinical, AS specific indexes, radiographic and biologic parameters were determined. HLA-B and B27 subtypes typing of all subjects were performed by PCR-SSP. Patients mean age was 38.4 years +/- 12.6 HLA-B27 was found in 62% of patients. The comparison of B27 positive and B27 negative patients revealed a correlation of B27 with age, male gender, family history of spondylarthropathies, age at disease onset, acute onset of the disease, inaugural spinal involvement, uveitis, bilateral and destructive hip arthritis as well as a high score of mSASSS. The most frequent sub-types of HLA B27 were B*2702 (49.2%) and B*2705 (36.3%). No significant difference of the clinical presentation of the disease or severity factors was found among these patients. This study confirmed the contribution of the HLA B27 to the determination of the clinical presentation of AS. The variability of factors linked to B27 may be explained by the polygenic model of the disease. PMID:22984764

Hamdi, W; Kaffel, D; Ghannouchi, M M; Laadhar, L; Makni, S; Kchir, M Montacer

2012-01-01

51

Different Subsets of Enteric Bacteria Induce and Perpetuate Experimental Colitis in Rats and Mice  

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Resident bacteria are incriminated in the pathogenesis of experimental colitis and inflammatory bowel diseases. We investigated the relative roles of various enteric bacteria populations in the induction and perpetuation of experimental colitis. HLA-B27 transgenic rats received antibiotics (ciprofloxacin, metronidazole, or vancomycin-imipenem) in drinking water or water alone in either prevention or treatment protocols. Mice were treated similarly with metronidazole or vancomycin-imipenem bef...

Rath, Heiko C.; Schultz, Michael; Freitag, Rene?; Dieleman, Levinus A.; Li, Fengling; Linde, Hans-jo?rg; Scho?lmerich, Ju?rgen; Sartor, R. Balfour

2001-01-01

52

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

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Natural HLA-B*2705 protein ligands with glutamine as anchor motif: implications for HLA-B27 association with spondyloarthropathy.  

Science.gov (United States)

The presentation of short viral peptide antigens by human leukocyte antigen (HLA) class I molecules on cell surfaces is a key step in the activation of cytotoxic T lymphocytes, which mediate the killing of pathogen-infected cells or initiate autoimmune tissue damage. HLA-B27 is a well known class I molecule that is used to study both facets of the cellular immune response. Using mass spectrometry analysis of complex HLA-bound peptide pools isolated from large amounts of HLA-B*2705(+) cells, we identified 200 naturally processed HLA-B*2705 ligands. Our analyses revealed that a change in the position (P) 2 anchor motif was detected in the 3% of HLA-B*2705 ligands identified. B*2705 class I molecules were able to bind these six GlnP2 peptides, which showed significant homology to pathogenic bacterial sequences, with a broad range of affinities. One of these ligands was able to bind with distinct conformations to HLA-B27 subtypes differentially associated with ankylosing spondylitis. These conformational differences could be sufficient to initiate autoimmune damage in patients with ankylosing spondylitis-associated subtypes. Therefore, these kinds of peptides (short, with GlnP2, and similar low affinity to all HLA-B27 subtypes tested but with unlike conformations in differentially ankylosing spondylitis-associated subtypes) must not be excluded from future researches involving potential arthritogenic peptides. PMID:23430249

Infantes, Susana; Lorente, Elena; Barnea, Eilon; Beer, Ilan; Barriga, Alejandro; Lasala, Fátima; Jiménez, Mercedes; Admon, Arie; López, Daniel

2013-04-12

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

Science.gov (United States)

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 diagnosis of inflammatory bowel disease difficult.

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

2014-01-01

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Multiple, non-conserved, internal viral ligands naturally presented by HLA-B27 in human respiratory syncytial virus-infected cells.  

Science.gov (United States)

Cytotoxic T lymphocyte (CTL)-mediated death of virus-infected cells requires prior recognition of short viral peptide antigens that are presented by human leukocyte antigen (HLA) class I molecules on the surface of infected cells. The CTL response is critical for the clearance of human respiratory syncytial virus (HRSV) infection. Using mass spectrometry analysis of complex HLA-bound peptide pools isolated from large amounts of HRSV-infected cells, we identified nine naturally processed HLA-B27 ligands. The isolated peptides are derived from six internal, not envelope, proteins of the infective virus. The sequences of most of these ligands are not conserved between different HRSV strains, suggesting a mechanism to explain recurrent infection with virus of different HRSV antigenic subgroups. In addition, these nine ligands represent a significant fraction of the proteome of this virus, which is monitored by the same HLA class I allele. These data have implications for vaccine development as well as for analysis of the CTL response. PMID:20081153

Infantes, Susana; Lorente, Elena; Barnea, Eilon; Beer, Ilan; Cragnolini, Juan José; García, Ruth; Lasala, Fátima; Jiménez, Mercedes; Admon, Arie; López, Daniel

2010-07-01

56

Multiple, Non-conserved, Internal Viral Ligands Naturally Presented by HLA-B27 in Human Respiratory Syncytial Virus-infected Cells*  

Science.gov (United States)

Cytotoxic T lymphocyte (CTL)-mediated death of virus-infected cells requires prior recognition of short viral peptide antigens that are presented by human leukocyte antigen (HLA) class I molecules on the surface of infected cells. The CTL response is critical for the clearance of human respiratory syncytial virus (HRSV) infection. Using mass spectrometry analysis of complex HLA-bound peptide pools isolated from large amounts of HRSV-infected cells, we identified nine naturally processed HLA-B27 ligands. The isolated peptides are derived from six internal, not envelope, proteins of the infective virus. The sequences of most of these ligands are not conserved between different HRSV strains, suggesting a mechanism to explain recurrent infection with virus of different HRSV antigenic subgroups. In addition, these nine ligands represent a significant fraction of the proteome of this virus, which is monitored by the same HLA class I allele. These data have implications for vaccine development as well as for analysis of the CTL response.

Infantes, Susana; Lorente, Elena; Barnea, Eilon; Beer, Ilan; Cragnolini, Juan Jose; Garcia, Ruth; Lasala, Fatima; Jimenez, Mercedes; Admon, Arie; Lopez, Daniel

2010-01-01

57

HLA-B27 Test  

Science.gov (United States)

... Association of America Arthritis Foundation American College of Rheumatology: Heredity and Arthritis Genetics Home Reference: HLA-B » ... Spondylitis and Inflammatory Bowel Disease. American College of Rheumatology from Arthritis & Rheumatism 2007; (DOI:10.1002/art. ...

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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

59

HLA B27 related 'unclassifiable' seronegative spondyloarthropathies.  

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Twenty-five patients (22 males and 3 females) are described who had 'unclassifiable' seronegative peripheral arthritis affecting mainly the large joints of the lower limbs with other typical features of spondyloarthropathies such as heel pain, low back pain, and mucosal ulcers. But their disorders could not be diagnosed as any specific spondyloarthropathy such as ankylosing spondylitis, Reiter's disease, etc. The mean age of onset of disease was 21.4 years and 60% of them had mono- or oligoar...

Prakash, S.; Mehra, N. K.; Bhargava, S.; Malaviya, A. N.

1983-01-01

60

Transgenic rat models of vasopressin overexpression.  

Science.gov (United States)

Vasopressin has an important role in water metabolism and its impairment induces some clinical disorders such as diabetes insipidus or syndrome of inappropriate antidiuresis (SIAD). SIAD is caused by the overproduction of vasopressin which induces diluting hyponatremia. The accurate diagnosis and appropriate therapy have not settled up to date because its pathophysiology is very complicated. It is meaningful to develop a rat model of SIAD in which human vasopressin gene is overexpressed in order to analyze pathophysiological changes. Several models transgenic for vasopressin including us had been generated. The transgenic rats provide a useful model to investigate various pathophysiological changes resulting from the oversecretion of vasopressin. Some interesting results based on these animal models are reviewed. PMID:14727685

Oiso, Yutaka; Nagasaki, Hiroshi; Yokoi, Hisashi

2003-11-01

 
 
 
 
61

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 ...

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

2008-01-01

62

Skeletal and cardiac myopathy in HIV-1 transgenic rats  

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The mechanism by which human immunodeficiency virus (HIV)-1 infection in humans leads to the erosion of lean body mass is poorly defined. Therefore, the purpose of the present study was to determine whether transgenic (Tg) rats that constitutively overexpress HIV-1 viral proteins exhibit muscle wasting and to elucidate putative mechanisms. Over 7 mo, Tg rats gained less body weight than pair-fed controls exclusively as a result of a proportional reduction in lean, not fat, mass. Fast- and slo...

Pruznak, Anne M.; Hong-brown, Ly; Lantry, Rachel; She, Pengxiang; Frost, Robert A.; Vary, Thomas C.; Lang, Charles H.

2008-01-01

63

Vascular damage without hypertension in transgenic rats expressing prorenin exclusively in the liver.  

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We have developed a transgenic animal model to investigate the effects of overexpression of rat prorenin on the cardiovascular system. Two transgenic rat lines were generated in which rat prorenin expression was directed to the liver by a human alpha1-antitrypsin promoter. Liver-specific expression was confirmed by RNase protection assay. Plasma prorenin concentrations in transgenic rats were increased 400-fold in the males of both lines but were increased only two- to threefold in the female...

Ve?niant, M.; Me?nard, J.; Bruneval, P.; Morley, S.; Gonzales, M. F.; Mullins, J.

1996-01-01

64

Immunodeficient Parameters in the HIV-1 Transgenic Rat Model  

Directory of Open Access Journals (Sweden)

Full Text Available Recently an HIV-1 transgenic (HIV-1Tg rat model was created that carries a gag-pol-deleted HIV-1 genome under the control of the HIV-1 viral promoter. However, other viral proteins are expressed in most organs and tissues, and are found in the circulating blood. Since HIV-1 targets the immune system in humans, we examined two immunological parameters, leukocyte-endothelial adhesion (LEA and inflammatory cytokine production, in 5 mo old HIV-1Tg rats to identify immune functions that may be impaired even before the onset of symptoms of HIV-1 infection. We administered a single injection (i.p. of the bacterial endotoxin, lipopolysaccharide (LPS, 250 ug/kg, to 5 mo old HIV-1Tg rats, age-matched transgenic control (Tg rats, and F344/NHsd (F344 control background strain rats. LPS induced an LEA response in both the Tg control and F344 control animals. However, in the HIV-1Tg rats, there was no LEA response to LPS. Following LPS administration, there was significantly greater serum levels of TNF-? and IL-1?, two pro-inflammatory cytokines, in the HIV-1Tg rats compared to the control animals. In contrast, the serum level of IL-10, an anti-inflammatory cytokine, was comparable in the HIV-1Tg, Tg control, and F344 control rats. Our data show that, in the HIV-1Tg rat, there is a negative correlation between the LEA response and the induction of pro-inflammatory cytokines in response to bacterial endotoxin. These findings suggest that the persistent presence of viral proteins may be, at least, partially responsible for the immunodeficiency that occurs with HIV-1 infection, and that the HIV-1Tg rat could be a valid rodent model in which to study various aspects of HIV-1 infection.

Sulie L. Chang

2007-01-01

65

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

2006-03-01

66

Activation of polyamine catabolism in transgenic rats induces acute pancreatitis  

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Polyamines are required for optimal growth and function of cells. Regulation of their cellular homeostasis is therefore tightly controlled. The key regulatory enzyme for polyamine catabolism is the spermidine/spermine N1-acetyltransferase (SSAT). Depletion of cellular polyamines has been associated with inhibition of growth and programmed cell death. To investigate the physiological function SSAT, we generated a transgenic rat line overexpressing the SSAT gene under the control of the inducib...

Alhonen, Leena; Parkkinen, Jyrki J.; Keina?nen, Tuomo; Sinervirta, Riitta; Herzig, Karl-heinz; Ja?nne, Juhani

2000-01-01

67

Focal cerebral ischemia in the TNFalpha-transgenic rat  

Directory of Open Access Journals (Sweden)

Full Text Available 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 infarct volume was calculated 24 hours or 7 days after one hour of reversible middle cerebral artery occlusion (MCAO. Cortical perfusion was monitored by laser-Doppler flowmetry (LDF during MCAO. Cortical vascular density was quantified by stereology. Post-ischemic cell death was assessed by immunohistochemistry and regional measurement of caspase-3 activity or DNA fragmentation. Unpaired t tests or analysis of variance with post hoc tests were used for comparison of group means. Results In TNF?-Tg rat brain, the aggregate mouse and rat TNF? mRNA level was fourfold higher than in non-Tg littermates and the corresponding TNF? protein level was increased fivefold (p ? 0.01. Infarct volume was greater in TNF?-Tg rats than in non-Tg controls at 24 hours (p ? 0.05 and 7 days (p ? 0.01. Within the first 10 minutes of MCAO, cortical perfusion measured by LDF was reduced in TNF?-Tg rats (p ? 0.05. However, regional vascular density was equivalent between TNF?-Tg and non-Tg animals (p = NS. Neural cellular apoptosis was increased in transgenic animals as shown by elevated caspase-3 activity (p ? 0.05 and DNA fragmentation (p ? 0.001 at 24 hours. Conclusion Chronic elevation of TNF? protein in brain increases susceptibility to ischemic injury but has no effect on vascular density. TNF?-Tg animals are more susceptible to apoptotic cell death after MCAO than are non-Tg animals. We conclude that the TNF?-Tg rat is a valuable new tool for the study of cytokine-mediated ischemic brain injury.

Springer Joe E

2008-10-01

68

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

2004-03-30

69

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

70

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

71

Transgenic rats with green, red, and blue fluorescence: powerful tools for bioimaging, cell trafficking, and differentiation  

Science.gov (United States)

The rat represents a perfect animal for broadening medical experiments, because its physiology has been well understood in the history of experimental animals. In addition, its larger body size takes enough advantage for surgical manipulation, compared to the mouse. Many rat models mimicking human diseases, therefore, have been used in a variety of biomedical studies including physiology, pharmacology, transplantation, and immunology. In an effort to create the specifically designed rats for biomedical research and regenerative medicine, we have developed the engineered rat system on the basis of transgenic technology and succeeded in establishing various transgenic rat strains. The transgenic rats with green fluorescent protein (GFP) were generated in the two different strains (Wistar and Lewis), in which GFP is driven under the chicken beta-actin promoter and cytomegalovirus enhancer (CAG promoter). Their GFP expression levels were different in each organ, but the Lewis line expressed GFP strongly and ubiquitously in most of the organs compared with that of Wistar. For red fluorescence, DsRed2 was transduced to the Wistar rats: one line specifically expresses DsRed2 in the liver under the mouse albumin promoter, another is designed for the Cre/LoxP system as the double reporter rat (the initial DsRed2 expression turns on GFP in the presence of Cre recombinase). LacZ-transgenic rats represent blue color, and LacZ is driven the CAG (DA) or ROSA26 promoter (Lewis). Our unique transgenic rats" system highlights the powerful performance for the elucidation of many cellular processes in regenerative medicine, leading to innovative medical treatments.

Murakami, Takashi; Kobayashi, Eiji

2005-04-01

72

Generation and characterization of a Tet-On (rtTA-M2 transgenic rat  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background The tetracycline-inducible gene regulation system is a powerful tool that allows temporal and dose-dependent regulation of target transgene expression in vitro and in vivo. Several tetracycline-inducible transgenic mouse models have been described with ubiquitous or tissue-specific expression of tetracycline-transactivator (tTA, reverse tetracycline-transactivator (rtTA or Tet repressor (TetR. Here we describe a Tet-On transgenic rat that ubiquitously expresses rtTA-M2 driven by the murine ROSA 26 promoter. Results The homozygous rat line (ROSA-rtTA-M2 generated by lentiviral vector injection, has a single integration site and was derived from the offspring of a genetic mosaic founder with multiple transgene integrations. The rtTA-M2 transgene integrated into an intron of a putative gene on chromosome 2 and does not appear to affect the tissue-specificity or expression of that gene. Fibroblasts from the ROSA-rtTA-M2 rats were transduced with a TetO7/CMV-EGFP lentivirus and exhibited doxycycline dose-dependent expression of the EGFP reporter transgene, in vitro. In addition, doxycycline-inducible EGFP expression was observed, in vivo, when the TetO7/CMV-EGFP lentivirus was injected into testis, kidney and muscle tissues of ROSA-rtTA-M2 rats. Conclusions This conditional expression rat model may have application for transgenic overexpression or knockdown studies of gene function in development, disease and gene therapy.

Sukhwani Meena

2010-02-01

73

Production of fat-1 transgenic rats using a post-natal female germline stem cell line.  

Science.gov (United States)

Germline stem cell lines possess the abilities of self-renewal and differentiation, and have been established from both mouse and human ovaries. Here, we established a new female germline stem cell (FGSC) line from post-natal rats by immunomagnetic sorting for Fragilis, which showed a normal karyotype, high telomerase activity, and a consistent gene expression pattern of primordial germ cells after 1 year of culture. Using an in vitro differentiation system, the FGSC line could differentiate into oocytes. After liposome-based transfection with green fluorescent protein (GFP) or fat-1 vectors, the FGSCs were transplanted into the ovaries of infertile rats. The transplanted FGSCs underwent oogenesis, and the rats produced offspring carrying the GFP or fat-1 transgene after mating with wild-type male rats. The efficiency of gene transfer was 27.86-28.00%, and 2 months was needed to produce transgenic rats. These findings have implications in biomedical research and potential applications in biotechnology. PMID:24258451

Zhou, Li; Wang, Lei; Kang, Jing X; Xie, Wenhai; Li, Xiaoyong; Wu, Changqing; Xu, Bo; Wu, Ji

2014-03-01

74

Development of transgenic rats producing human ?-amyloid precursor protein as a model for Alzheimer's disease: Transgene and endogenous APP genes are regulated tissue-specifically  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Alzheimer's disease (AD is a devastating neurodegenerative disorder that affects a large and growing number of elderly individuals. In addition to idiopathic disease, AD is also associated with autosomal dominant inheritance, which causes a familial form of AD (FAD. Some instances of FAD have been linked to mutations in the ?-amyloid protein precursor (APP. Although there are numerous mouse AD models available, few rat AD models, which have several advantages over mice, have been generated. Results Fischer 344 rats expressing human APP driven by the ubiquitin-C promoter were generated via lentiviral vector infection of Fischer 344 zygotes. We generated two separate APP-transgenic rat lines, APP21 and APP31. Serum levels of human amyloid-beta (A?40 were 298 pg/ml for hemizygous and 486 pg/ml for homozygous APP21 animals. Serum A?42 levels in APP21 homozygous rats were 135 pg/ml. Immunohistochemistry in brain showed that the human APP transgene was expressed in neurons, but not in glial cells. These findings were consistent with independent examination of enhanced green fluorescent protein (eGFP in the brains of eGFP-transgenic rats. APP21 and APP31 rats expressed 7.5- and 3-times more APP mRNA, respectively, than did wild-type rats. Northern blots showed that the human APP transgene, driven by the ubiquitin-C promoter, is expressed significantly more in brain, kidney and lung compared to heart and liver. A similar expression pattern was also seen for the endogenous rat APP. The unexpected similarity in the tissue-specific expression patterns of endogenous rat APP and transgenic human APP mRNAs suggests regulatory elements within the cDNA sequence of APP. Conclusion This manuscript describes the generation of APP-transgenic inbred Fischer 344 rats. These are the first human AD model rat lines generated by lentiviral infection. The APP21 rat line expresses high levels of human APP and could be a useful model for AD. Tissue-specific expression in the two transgenic rat lines and in wild-type rats contradicts our current understanding of APP gene regulation. Determination of the elements that are responsible for tissue-specific expression of APP may enable new treatment options for AD.

Chan Anthony WS

2008-02-01

75

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

76

Overexpression of vasopressin in the rat transgenic for the metallothionein-vasopressin fusion gene.  

Science.gov (United States)

Arginine vasopressin (AVP) is a major antidiuretic hormone, the overproduction of which causes diluting hyponatremia in humans and is called the syndrome of inappropriate antidiuresis (SIAD). To study physiological changes resulting from AVP overproduction and to develop an animal model of hyponatremia, the human AVP gene was expressed under the control of the metallothionein promoter in transgenic (Tg) rats. Analyses of AVP immunoreactivity (irAVP) in the tissues revealed that the transgene is expressed mainly in the central nervous system. Gel filtration showed that irAVP in the brain and plasma was properly processed AVP. AVP purified from the brains of both Tg and control rats also exerted equal bioactivity to generate cAMP in LLC-PK1 cells. The founder rats did not show any physical or anatomical abnormalities. Under basal conditions, Tg rats had high plasma AVP levels (Tg 13.8 +/- 2.5 pg/ml; control 2.7 +/- 1.2 pg/ml; n=6 in both groups; means +/- S.E.M.), decreased urine volume, and normal plasma [Na(+)]. Hypertonic saline injected i.p. did not affect AVP secretion in Tg rats. In response to a zinc-supplemented liquid diet, plasma AVP decreased in control rats, but increased in Tg rats (Tg 32.7 +/- 2.7 pg/ml; control 1.0+/-0.1 pg/ml; n=6), resulting in hyponatremia (Tg 135.2 +/- 2.5 mEq/l; control 140.8 +/- 0.4 mEq/l; n=6). To our knowledge, this is the first transgenic animal to show diluting hyponatremia. This transgenic rat may therefore provide a useful model in which to investigate various physiological alterations resulting from the oversecretion of AVP which involve SIAD, stress response, behavior, and blood pressure. PMID:11927382

Nagasaki, H; Yokoi, H; Arima, H; Hirabayashi, M; Ishizaki, S; Tachikawa, K; Murase, T; Miura, Y; Oiso, Y

2002-04-01

77

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

78

Development of Hyperplasias, Preneoplasias, and Mammary Tumors in MMTV-c-erbB-2 and MMTV-TGF? Transgenic Rats  

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Human cDNAs corresponding to two epidermal growth factor-related products that are overexpressed in human breast cancers, that for c-erbB-2 (HER-2) and for transforming growth factor ? (TGF?), have been cloned downstream of the mouse mammary tumor virus (MMTV) long terminal repeat promoter and injected into the pronucleus of fertilized oocytes of Sprague-Dawley rats to produce transgenic offspring. Expression of the transgenic mRNAs is not detectable in mammary tissue from virgin transgenic...

Davies, Barry R.; Platt-higgins, Angela M.; Schmidt, Gunter; Rudland, Philip S.

1999-01-01

79

Interaction between ERAP1 and HLA-B27 in ankylosing spondylitis implicates peptide handling in the mechanism for HLA-B27 in disease susceptibility  

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Ankylosing spondylitis is a common form of inflammatory arthritis predominantly affecting the spine and pelvis that occurs in approximately 5 out of 1,000 adults of European descent. Here we report the identification of three variants in the RUNX3, LTBRTNFRSF1A and IL12B regions convincingly associated with ankylosing spondylitis (P < 5 × 10?8 in the combined discovery and replication datasets) and a further four loci at PTGER4, TBKBP1, ANTXR2 and CARD9 that show strong association across ...

Evans, David M.; Spencer, Chris C. A.; Pointon, Jennifer J.; Su, Zhan; Harvey, David; Kochan, Grazyna; Oppermann, Udo; Dilthey, Alexander; Pirinen, Matti; Stone, Millicent A.; Appleton, Louise; Moutsianas, Loukas; Leslie, Stephen; Wordsworth, Tom; Kenna, Tony J.

2011-01-01

80

Transgenic Rat Model of Neurodegeneration Caused by Mutation in the TDP Gene  

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TDP-43 proteinopathies have been observed in a wide range of neurodegenerative diseases. Mutations in the gene encoding TDP-43 (i.e., TDP) have been identified in amyotrophic lateral sclerosis (ALS) and in frontotemporal lobe degeneration associated with motor neuron disease. To study the consequences of TDP mutation in an intact system, we created transgenic rats expressing normal human TDP or a mutant form of human TDP with a M337V substitution. Overexpression of mutant, but not normal, TDP...

Zhou, Hongxia; Huang, Cao; Chen, Han; Wang, Dian; Landel, Carlisle P.; Xia, Pedro Yuxing; Bowser, Robert; Liu, Yong-jian; Xia, Xu Gang

2010-01-01

 
 
 
 
81

Retinal Degeneration in Two Lines of Transgenic S334ter Rats  

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Aim of this study was to examine synaptic connectivity changes in the retina and the location and rate of apoptosis in transgenic S334ter line-3 and line-5 rats with photoreceptor degeneration. Heterozygous S334ter-line-3 and line-5 at P11-13, P30, P60, P90 and several control non-dystrophic rats (Long Evans and Sprague-Dawley) at P60, were studied anatomically by immunohistochemistry for various cell and synaptic markers, and by PNA and TUNEL label.- S334ter line-3 exhibited the fastest rate...

Martinez-navarrete, G.; Seiler, M. J.; Aramant, R. B.; Fernandez-sanchez, L.; Pinilla, I.; Cuenca, N.

2011-01-01

82

Profiles of motor and cognitive impairment in the transgenic rat model of Huntington's disease.  

Science.gov (United States)

The transgenic Huntington's disease (tgHD) rat strain provides a well regarded transgenic animal model of Huntington's disease, offering the prospect for a more detailed functional analysis in rats, along with neurological and therapeutic interventions, than is possible in the more widely available mouse models. In the present experiments, we compare the performance of heterozygous and homozygous tgHD rats against wildtype littermates on a range of motor and cognitive assessments in five separate cohorts of rats between 8 and 22 months of age. Male but not female heterozygous tgHD rats exhibit modest motor deficits in rotarod and staircase reaching tests, whereas most cognitive tests (including object recognition, exploration of novelty, delayed alternation, choice reaction time, and serial implicit learning tasks) revealed at best small or inconsistent deficits, in homozygous as well as heterozygous animals, up to 22 months of age. Thus, although we have observed modest but clear-cut deficits in motor phenotype, with a sex difference in line with previous reports, we have not established a robust cognitive impairment in this strain on a range of tasks sensitive to frontostriatal function, as required for testing novel (symptomatic, protective or reparative) therapeutics in a robust, valid, animal model of human Huntington's disease. PMID:21963415

Fielding, Steven A; Brooks, Simon P; Klein, Alexander; Bayram-Weston, Zubeyde; Jones, Lesley; Dunnett, Stephen B

2012-06-01

83

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

Science.gov (United States)

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.

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

2014-01-01

84

Rosuvastatin ameliorates the development of pulmonary arterial hypertension in the transgenic (mRen2)27 rat  

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We have recently reported that transgenic (mRen2)27 rats (Ren2 rats) exhibit pulmonary arterial hypertension (PAH), which is, in part, mediated by oxidative stress. Since 3-hydroxy-3-methylglutaryl-CoA reductase inhibitors (statins) exhibit beneficial vascular effects independent of cholesterol synthesis, we hypothesized that rosuvastatin (RSV) treatment ameliorates PAH and pulmonary vascular remodeling in Ren2 rats, in part, by reducing oxidative stress. Six-week-old male Ren2 and Sprague-Da...

2009-01-01

85

HTLV-I env-pX transgenic rats: prototype animal model for collagen vascular diseases.  

Science.gov (United States)

To evaluate the function of HTLV-I env-pX gene in vivo, we developed two lines of transgenic rats (env-pX rats) that expressed env-pX gene products, under control of own LTR promotor. In various tissues of the rats, env and pX mRNAs were constitutively expressed, irrespective of age. At age 5 weeks, swelling of the bilateral ankle joints histologically showing synovial lining hyperplasia, severe chronic inflammation, erosion of the joint cartilage, and bone destruction with pannus formation began to develop in these env-pX rats. These histologic features resemble those of rheumatoid arthritis (RA) in man. High titered rheumatoid factors and low anti-dsDNA antibodies and hyper-gamma globulinemia were detected. Necrotizing arteritis resembling polyarteritis nodosa, polymyositis, myocarditis and Sjögren syndrome-like sialoadenitis developed, together with RA-like arthritis even in one individual animal. Thymic atrophy with low body weight was also observed. The evidence indicates that env-pX rats appear to be suitable animal models for elucidating pathogenetic mechanisms involved in not only HTLV-I related diseases but also various collegen vascular and autoimmune diseases of unknown etiology in man. PMID:9209358

Yamazaki, H; Ikeda, H; Ishuzu, A; Shikishima, H; Kikuchi, K; Wakisaka, A; Hatanaka, M; Yoshiki, T

1997-04-01

86

Radiographic visualisation of seropositive rheumatoid arthritis in Carriers of HLA-B27  

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A group of 11 B27-positive, seropositive patients with rheumatoid arthritis was compared with 11 matched B27-negative seropositive patients. The radiographs of all limb joints, the sacroiliac joints, and the cervical spine were read blindly. Ten patients in each group were radiographed 2-6 times during observation periods of 3-13 years; one patient in each group was only examined once. The prevailing picture of both groups was that of progressive erosive rheumatoid arthritis, although two small differences were found: Erosions of the apophyseal joints of the cervical spine and slight periosteal new bone formation of the shoulder, hip, and knee regions occurred more often in the B27-positive than in the B27-negative group.

Jurik, A.G.; Carvalho, A. de; Graudal, H.

1987-07-01

87

Radiographic visualisation of seropositive rheumatoid arthritis in Carriers of HLA-B27  

International Nuclear Information System (INIS)

A group of 11 B27-positive, seropositive patients with rheumatoid arthritis was compared with 11 matched B27-negative seropositive patients. The radiographs of all limb joints, the sacroiliac joints, and the cervical spine were read blindly. Ten patients in each group were radiographed 2-6 times during observation periods of 3-13 years; one patient in each group was only examined once. The prevailing picture of both groups was that of progressive erosive rheumatoid arthritis, although two small differences were found: Erosions of the apophyseal joints of the cervical spine and slight periosteal new bone formation of the shoulder, hip, and knee regions occurred more often in the B27-positive than in the B27-negative group. (orig.)

1987-01-01

88

Endothelial dysfunction in the aorta of transgenic rats harboring the mouse Ren-2 gene.  

Science.gov (United States)

The renin-angiotensin system plays an important role in the pathophysiology of hypertension. We studied vascular function in the aorta of mouse Ren-2 transgenic rats (TGR(mRen2)27). Changes in isometric tension of isolated aorta of TGR(mRen2)27 and Sprague-Dawley rats (SD) were recorded in organ chambers. Contractions to angiotensin II (AII), big-endothelin and endothelin-1 (ET-1), but not KCl were decreased in TGR. Blockade of nitric oxide (NO)-synthase by L-NAME or removal of the endothelium did not alter these decreased contractions to ET-1 and AII in TGR, suggesting that receptors or signaling pathways of these two agonists are downregulated during hypertension. Contractions to norepinephrine (NE) were also lower in TGR, however blockade of NO-synthase by L-NAME or removal of the endothelium evoked similar contractions to NE in both strains, suggesting that basal release of NO reduces contractions to NE to a greater extent in transgenic than control rats. In the presence of L-NAME, acetylcholine evoked endothelium-dependent contractions (EDCF) in TGR, which were blocked by the thromboxane/prostaglandin H2 receptor antagonists SQ 30741, and partially by the thromboxane synthase inhibitor CGS 13080, suggesting that prostaglandin H2 is the mediator. Endothelium-dependent relaxation to acetylcholine was decreased in TGR, while endothelium-independent relaxations to sodium nitroprusside were similar in both strains. SQ 30741 did not improve relaxations to acetylcholine in TGR indicating that impaired relaxations to acetylcholine are due to a decreased acetylcholine-receptor mediated release of NO rather than increased release of EDCF. Thus, Ren-2 hypertension leads to marked alterations of vascular functions in the aorta. These changes could contribute to hypertension and its vascular complications in TGR(mRen2)27 rats. PMID:10365769

Arnet, U A; Novosel, D; Barton, M; Noll, G; Ganten, D; Lüscher, T F

1999-01-01

89

MUTAGENICITY OF 3-METHYLCHOLANTHRENE, PCB3, AND 4-OH-PCB3 IN THE LUNG OF TRANSGENIC BIGBLUE® RATS  

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Recent findings of high levels of predominantly lower chlorinated biphenyls in indoor and outdoor air open the question of possible health consequences. Lower chlorinated biphenyls are more readily metabolized to reactive and potentially harmful intermediates, acting as mutagens and cancer initiators. The goal of this study was to assess the mutagenicity of PCB3 in the lungs of rats. Male BigBlue® 334 Fisher transgenic rats, which carry the bacterial lacI gene as a target of mutagenicity, we...

Maddox, Catherine; Wang, Bingxuan; Kirby, Patricia A.; Wang, Kai; Ludewig, Gabriele

2008-01-01

90

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

91

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

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Correction to Rao J S, Kim H W, Kellom M, Greenstein D, Chen M, Kraft A D, Harry G J, Rapoport S I, Basselin M. Increased neuroinflammatory and arachidonic acid cascade markers, and reduced synaptic proteins, in brain of HIV-1 transgenic rats. Journal of Neuroinflammation 8:101.

Rao Jagadeesh

2012-01-01

92

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

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Correction to Rao J S, Kim H W, Kellom M, Greenstein D, Chen M, Kraft A D, Harry G J, Rapoport S I, Basselin M. Increased neuroinflammatory and arachidonic acid cascade markers, and reduced synaptic proteins, in brain of HIV-1 transgenic rats. Journal of Neuroinflammation 8:101.

Rao, Jagadeesh Sridhara; Kim, Hyung-wook; Kellom, Matthew; Greenstein, Dede; Chen, Mei; Kraft, Andrew David; Harry, Gaylia Jean; Rapoport, Stanley Isaac; Basselin, Mireille

2012-01-01

93

A transgenic Alzheimer rat with plaques, tau pathology, behavioral impairment, oligomeric A? and frank neuronal loss  

Science.gov (United States)

Alzheimer’s disease (AD) is hallmarked by amyloid plaques, neurofibrillary tangles, and widespread cortical neuronal loss (Selkoe, 2001). The ‘amyloid cascade hypothesis’ posits that cerebral amyloid sets neurotoxic events into motion that precipitate Alzheimer dementia (Hardy and Allsop, 1991). Yet, faithful recapitulation of all AD features in widely used transgenic (Tg) mice engineered to overproduce A? peptides has been elusive. We have developed a Tg rat model (line TgF344-AD) expressing mutant human amyloid precursor protein (APPsw) and presenilin 1 (PS1?E9) genes, each independent causes of early-onset familial AD. TgF344-AD rats manifest age-dependent cerebral amyloidosis that precedes tauopathy, gliosis, apoptotic loss of neurons in the cerebral cortex and hippocampus, and cognitive disturbance. These results demonstrate progressive neurodegeneration of the Alzheimer type in these animals. The TgF344-AD rat fills a critical need for a next-generation animal model to enable basic and translational AD research.

Cohen, Robert M.; Rezai-Zadeh, Kavon; Weitz, Tara M.; Rentsendorj, Altan; Gate, David; Spivak, Inna; Bholat, Yasmin; Vasilevko, Vitaly; Glabe, Charles G.; Breunig, Joshua J.; Rakic, Pasko; Davtyan, Hayk; Agadjanyan, Michael G.; Kepe, Vladimir; Barrio, Jorge; Bannykh, Serguei; Szekely, Christine A.; Pechnick, Robert N.; Town, Terrence

2013-01-01

94

Expression of ATP-binding cassette membrane transporters in a HIV-1 transgenic rat model.  

Science.gov (United States)

P-glycoprotein (P-gp, product of Mdr1a and Mdr1b genes), multidrug resistance associated proteins (Mrps), and breast cancer resistance protein (Bcrp), all members of the ATP-binding cassette (ABC) membrane-associated drug transporters superfamily, can significantly restrict the entry of antiretroviral drugs (ARVs) into organs which exhibit a barrier function such as the central nervous system (CNS) and the male genital tract (MGT). In vitro, HIV-1 viral proteins such as glycoprotein-120 (gp120) and transcriptional transactivator (tat) have been shown to alter the expression of these transporters and ARVs permeability. The objective of this study was to compare mRNA expression of these transporters, in vivo, in several tissues obtained from HIV-1 transgenic rats (Tg-rat) (8 and 24 weeks) with those of age-matched wild-type rats. At 24 weeks, significant changes in several drug transporter mRNA expressions were observed, in particular, in brain, kidney, liver and testes. These findings suggest that HIV-1 viral proteins can alter the expression of ABC drug transporters, in vivo, in the context of HIV-1 and further regulate ARVs permeability in several organs including the CNS and MGT, two sites which have been reported to display very low ARVs permeability in the clinic. PMID:24472536

Robillard, Kevin R; Hoque, Md Tozammel; Bendayan, Reina

2014-02-21

95

Ultrastructure Study of Transgenic Ren2 Rat Aorta - Part 1: Endothelium and Intima  

Science.gov (United States)

Background The renin-angiotensin-aldosterone system plays an important role in the development and progression of hypertension and accelerated atherosclerosis (atheroscleropathy) associated with the cardiorenal metabolic syndrome and type 2 diabetes mellitus. Additionally, the renin-angiotensin-aldosterone system plays an important role in vascular-endothelial-intimal cellular and extracellular remodeling. Methods Thoracic aortas of young male transgenic heterozygous (mRen2)27 (Ren2) rats were utilized for this ultrastructural study. This lean model of hypertension, insulin resistance and oxidative stress harbors the mouse renin gene with increased local tissue (aortic) levels of angiotensin II and angiotensin type 1 receptors and elevated plasma aldosterone levels. Results The ultrastructural observations included marked endothelial cell retraction, separation, terminal nuclear lifting, adjacent duplication, apoptosis and a suggestion of endothelial progenitor cell attachment. The endothelium demonstrated increased caveolae, microparticles, depletion of Weibel-Palade bodies, loss of cell-cell and basal adhesion hemidesmosome-like structures, platelet adhesion and genesis of subendothelial neointima. Conclusion These observational ultrastructural studies of the transgenic Ren2 vasculature provide an in-depth evaluation of early abnormal remodeling changes within conduit-elastic arteries under conditions of increased local levels of angiotensin II, oxidative stress, insulin resistance and hypertension.

Hayden, Melvin R.; Habibi, Javad; Joginpally, Tejaswini; Karuparthi, Poorna R.; Sowers, James R.

2012-01-01

96

Assessment of Motor Function, Sensory Motor Gating and Recognition Memory in a Novel BACHD Transgenic Rat Model for Huntington Disease  

Science.gov (United States)

Rationale Huntington disease (HD) is frequently first diagnosed by the appearance of motor symptoms; the diagnosis is subsequently confirmed by the presence of expanded CAG repeats (> 35) in the HUNTINGTIN (HTT) gene. A BACHD rat model for HD carrying the human full length mutated HTT with 97 CAG-CAA repeats has been established recently. Behavioral phenotyping of BACHD rats will help to determine the validity of this model and its potential use in preclinical drug discovery studies. Objectives The present study seeks to characterize the progressive emergence of motor, sensorimotor and cognitive deficits in BACHD rats. Materials and Methods Wild type and transgenic rats were tested from 1 till 12 months of age. Motor tests were selected to measure spontaneous locomotor activity (open field) and gait coordination. Sensorimotor gating was assessed in acoustic startle response paradigms and recognition memory was evaluated in an object recognition test. Results Transgenic rats showed hyperactivity at 1 month and hypoactivity starting at 4 months of age. Motor coordination imbalance in a Rotarod test was present at 2 months and gait abnormalities were seen in a Catwalk test at 12 months. Subtle sensorimotor changes were observed, whereas object recognition was unimpaired in BACHD rats up to 12 months of age. Conclusion The current BACHD rat model recapitulates certain symptoms from HD patients, especially the marked motor deficits. A subtle neuropsychological phenotype was found and further studies are needed to fully address the sensorimotor phenotype and the potential use of BACHD rats for drug discovery purposes.

Abada, Yah-se K.; Nguyen, Huu Phuc; Schreiber, Rudy; Ellenbroek, Bart

2013-01-01

97

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...

2007-01-01

98

Overexpression of the rat sarcoplasmic reticulum Ca2+ ATPase gene in the heart of transgenic mice accelerates calcium transients and cardiac relaxation.  

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The Ca2+ ATPase of the sarcoplasmic reticulum (SERCA2) plays a dominant role in lowering cytoplasmic calcium levels during cardiac relaxation and reduction of its activity has been linked to delayed diastolic relaxation in hypothyroid and failing hearts. To determine the contractile alterations resulting from increased SERCA2 expression, we generated transgenic mice overexpressing a rat SERCA2 transgene. Characterization of a heterozygous transgenic mouse line (CJ5) showed that the amount of ...

He, H.; Giordano, F. J.; Hilal-dandan, R.; Choi, D. J.; Rockman, H. A.; Mcdonough, P. M.; Bluhm, W. F.; Meyer, M.; Sayen, M. R.; Swanson, E.; Dillmann, W. H.

1997-01-01

99

Generation of topically transgenic rats by in utero electroporation and in vivo bioluminescence screening.  

Science.gov (United States)

In utero electroporation (IUE) is a technique which allows genetic modification of cells in the brain for investigating neuronal development. So far, the use of IUE for investigating behavior or neuropathology in the adult brain has been limited by insufficient methods for monitoring of IUE transfection success by non-invasive techniques in postnatal animals. For the present study, E16 rats were used for IUE. After intraventricular injection of the nucleic acids into the embryos, positioning of the tweezer electrodes was critical for targeting either the developing cortex or the hippocampus. Ventricular co-injection and electroporation of a luciferase gene allowed monitoring of the transfected cells postnatally after intraperitoneal luciferin injection in the anesthetized live P7 pup by in vivo bioluminescence, using an IVIS Spectrum device with 3D quantification software. Area definition by bioluminescence could clearly differentiate between cortical and hippocampal electroporations and detect a signal longitudinally over time up to 5 weeks after birth. This imaging technique allowed us to select pups with a sufficient number of transfected cells assumed necessary for triggering biological effects and, subsequently, to perform behavioral investigations at 3 month of age. As an example, this study demonstrates that IUE with the human full length DISC1 gene into the rat cortex led to amphetamine hypersensitivity. Co-transfected GFP could be detected in neurons by post mortem fluorescence microscopy in cryosections indicating gene expression present at ?6 months after birth. We conclude that postnatal bioluminescence imaging allows evaluating the success of transient transfections with IUE in rats. Investigations on the influence of topical gene manipulations during neurodevelopment on the adult brain and its connectivity are greatly facilitated. For many scientific questions, this technique can supplement or even replace the use of transgenic rats and provide a novel technology for behavioral neuroscience. PMID:24084570

Vomund, Sandra; Sapir, Tamar; Reiner, Orly; Silva, Maria A de Souza; Korth, Carsten

2013-01-01

100

Ribozyme rescue of photoreceptor cells in a transgenic rat model of autosomal dominant retinitis pigmentosa.  

Science.gov (United States)

Ribozymes, catalytic RNA molecules that cleave a complementary mRNA sequence, have potential as therapeutics for dominantly inherited disease. Twelve percent of American patients with the blinding disease autosomal dominant retinitis pigmentosa (ADRP) carry a substitution of histidine for proline at codon 23 (P23H) in their rhodopsin gene, resulting in photoreceptor cell death from the synthesis of the abnormal gene product. Ribozymes can discriminate and catalyze the in vitro destruction of P23H mutant mRNAs from a transgenic rat model of ADRP. Here, we demonstrate that in vivo expression of either a hammerhead or hairpin ribozyme in this rat model considerably slows the rate of photoreceptor degeneration for at least three months. Catalytically inactive control ribozymes had less effect on the retinal degeneration. Intracellular production of ribozymes in photoreceptors was achieved by transduction with a recombinant adeno-associated virus (rAAV) incorporating a rod opsin promoter. Ribozyme-directed cleavage of mutant mRNAs, therefore, may be an effective therapy for ADRP and also may be applicable to other inherited diseases. PMID:9701253

Lewin, A S; Drenser, K A; Hauswirth, W W; Nishikawa, S; Yasumura, D; Flannery, J G; LaVail, M M

1998-08-01

 
 
 
 
101

The Lewis GFP transgenic rat strain is a useful cell donor for neural transplantation.  

Science.gov (United States)

Stem cell transplantation is a promising therapeutic approach in neurodegenerative diseases. Studying graft survival and development has important implications for the further development of experimental and clinical transplantation protocols. Cellular elements in neural transplants are sometimes difficult to identify. The existing labeling methods cannot reliably provide stably labeled cells that can be detected in long-term experiments. Transgenic (tg) Lewis rats ubiquitously expressing green fluorescent protein (GFP) provide an ideal donor source. The aim of this project was to investigate the potential of GFP-tg Lewis rats to serve as donor tissue for neural stem cell transplantation. Ventral mesencephalon (VM) GFP-tg E14.5-derived cells were compared to wild-type (wt) in vitro and in vivo. Firstly, cells from GFP and non-GFP VM tissue were compared with regard to their proliferation and response towards 6-OHDA-toxicity in culture. Secondly, 6-OHDA-lesioned hemiparkinsonian Sprague-Dawley/Crl:CD(SD) rats received intrastriatal grafts derived from VM of E14.5 GFP-tg rats. Due to the fact that donor and recipient belong to two different rat strains, we focused on graft survival in correlation with immunosuppression and graft GFP and tyrosine hydroxylase (TH) expression. In summary, in vitro tg cells exhibited 98% GFP expression and did not differ from wt cells in any of the measured parameters. In vivo, all experimental groups showed a significant compensation in rotation behavior after transplantation. Furthermore, there was no difference on rotation behavior or graft morphology and survival pattern as well as GFP expression between immunosuppressed and nonimmunosuppressed animals. The GFP-positive population of the graft was composed of 13.3% GFAP-positive, 56.1% NeuN-positive, and 1.9% TH-positive cells. Analysis of graft subpopulations manifested that 70.6% of GFAP-positive, 86.9% of NeuN-positive, and 80.1% of TH-positive cells coexpressed GFP. In conclusion, our data show that the Lewis GFP-tg rats serve as an excellent cell source for studying primary neural precursor cells in the transplantation paradigm. PMID:22405077

Krause, Martin; Ganser, Claudia; Kobayashi, Eiji; Papazoglou, Anna; Nikkhah, Guido

2012-01-01

102

Estrogen receptor beta expression and apoptosis of spermatocytes of mice overexpressing a rat androgen-binding protein transgene.  

Science.gov (United States)

Progression of the first meiotic division in male germ cells is regulated by a variety of factors, including androgens and possibly estrogens. When this regulation fails, meiosis is arrested and primary spermatocytes degenerate by apoptosis. Earlier studies showed that overexpression of rat androgen-binding protein (ABP) in the testis of transgenic mice results in a partial meiotic arrest and apoptosis of pachytene spermatocytes. In view of the recent localization of estrogen receptor beta (ERbeta) in primary spermatocytes and data suggesting the ability of ERbeta to repress cellular proliferation, we tested the hypothesis that variations in the testicular steroid microenvironment caused by excess ABP produce changes in ERbeta expression in this cellular type that could be associated to the meiotic arrest and, eventually, to the induction of germ cell apoptosis observed in the ABP transgenic mice. Increased levels of ERbeta mRNA and protein were demonstrated in the testis of rat ABP transgenic mice compared with nontransgenic littermates by reverse transcriptase-polymerase chain reaction (RT-PCR) experiments, Northern blotting, and Western Blotting. The major differences were found when isolated germ cells of transgenic and nontransgenic littermates were analyzed by RT-PCR. In keeping with this finding, ERbeta was strongly immunolabeled in pachytene spermatocytes of rat ABP transgenic mice and localized in tubular stages in which TUNEL labeling was maximal. Confocal microscopy analysis of a fluorescent TUNEL assay and ERbeta immunohistochemistry revealed that degenerating pachytene spermatocytes overexpressed ERbeta. The present results are consistent with the interpretation that ERbeta is associated with the events that regulate negatively the progression of meiosis or that lead to spermatocyte apoptosis. PMID:15215204

Selva, David M; Tirado, Oscar M; Toràn, Nuria; Suárez-Quian, Carlos A; Reventos, Jaume; Munell, Francina

2004-11-01

103

Renin Inhibition Attenuates Insulin Resistance, Oxidative Stress, and Pancreatic Remodeling in the Transgenic Ren2 Rat  

Science.gov (United States)

Emerging evidence indicates that pancreatic tissue expresses all components of the renin-angiotensin system. However, the functional role is not well understood. This investigation examined renin inhibition on pancreas structure/function in the transgenic Ren2 rat harboring the mouse renin gene, a model of tissue renin overexpression. Renin is the rate-limiting step in the generation of angiotensin II (Ang II), which stimulates the generation of reactive oxygen species in a variety of tissues. Overexpression of renin in Ren2 rats results in hypertension, insulin resistance, and cardiovascular and renal damage. Young (6–7 wk old) insulin-resistant male Ren2 and age-matched insulin sensitive Sprague Dawley rats were treated with the renin inhibitor, aliskiren (50 mg/kg·d by ip injection), or placebo for 21 d. At 21 d, the Ren2 demonstrated insulin resistance with increased islet insulin, Ang II, and reduced total insulin receptor substrate (IRS)-1, IRS-2, and Akt immunostaining. There was increased islet nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity and subunits (p47phox and Rac1) as well as increased nitrotyrosine immunostaining (each P < 0.05). These functional abnormalities were associated with a disordered islet architecture; increased islet-exocrine interface, pericapillary fibrosis, and structurally abnormal mitochondria and content in endocrine and exocrine pancreas. In vivo treatment with aliskiren normalized systemic insulin resistance and islet insulin, Ang II, NADPH oxidase activity/subunits, and nitrotyrosine and improved total IRS-1 and Akt phosphorylation (each P < 0.05) as well as islet/exocrine structural abnormalities. Collectively, these data suggest that pancreatic functional/structural changes are driven, in part, by tissue renin-angiotensin system-mediated increases in NADPH oxidase and reactive oxygen species generation, abnormalities attenuated with direct renin inhibition.

Habibi, Javad; Whaley-Connell, Adam; Hayden, Melvin R.; DeMarco, Vincent G.; Schneider, Rebecca; Sowers, Susan D.; Karuparthi, Poorna; Ferrario, Carlos M.; Sowers, James R.

2008-01-01

104

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

International Nuclear Information System (INIS)

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

2005-05-13

105

Progress Toward a Human CD4/CCR5 Transgenic Rat Model for De Novo Infection by Human Immunodeficiency Virus Type 1  

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The development of a permissive small animal model for the study of human immunodeficiency virus type (HIV)-1 pathogenesis and the testing of antiviral strategies has been hampered by the inability of HIV-1 to infect primary rodent cells productively. In this study, we explored transgenic rats expressing the HIV-1 receptor complex as a susceptible host. Rats transgenic for human CD4 (hCD4) and the human chemokine receptor CCR5 (hCCR5) were generated that express the transgenes in CD4+ T lymph...

Keppler, Oliver T.; Welte, Frank J.; Ngo, Tuan A.; Chin, Peggy S.; Patton, Kathryn S.; Tsou, Chia-lin; Abbey, Nancy W.; Sharkey, Mark E.; Grant, Robert M.; You, Yun; Scarborough, John D.; Ellmeier, Wilfried; Littman, Dan R.; Stevenson, Mario; Charo, Israel F.

2002-01-01

106

3-Methylcholanthrene (3-MC) and 4-Chlorobiphenyl (PCB3) genotoxicity is gender-related in Fischer 344 transgenic rats  

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Polychlorinated biphenyls (PCBs) are a class of persistent organic pollutants with myriad biological effects, including carcinogenicity. We present data showing gender-specific genotoxicity in Fischer 344 transgenic BigBlue rodents exposed to 4-chlorobiphenyl (PCB3), a hydroxylated metabolite, and the positive control 3-methylcholanthrene (3-MC) where female rats are more resistant to the genotoxic effects of the test compounds compared to their male counterparts. This difference is further h...

Jacobus, J. A.; Wang, B.; Maddox, C.; Esch, H.; Lehmann, L.; Robertson, L. W.; Wang, K.; Kirby, P.; Ludewig, G.

2010-01-01

107

Comparative analysis of telmisartan and olmesartan on cardiac function in the transgenic (mRen2)27 rat  

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Telmisartan, an angiotensin receptor blocker, may have unique benefits as it possesses partial peroxisome proliferator-activated receptor (PPAR)-? agonist activity in addition to antihypertensive effects. In this study, we test whether treatment with telmisartan ameliorates cardiovascular abnormalities to a greater extent than olmesartan, which has little PPAR-? activity. The hypertensive rodent model of tissue renin-angiotensin system activation, transgenic (mRen2)27 (Ren2) rats and their ...

Demarco, Vincent G.; Johnson, Megan S.; Habibi, Javad; Pulakat, Lakshmi; Gul, Rukhsana; Hayden, Melvin R.; Tilmon, Roger D.; Dellsperger, Kevin C.; Winer, Nathaniel; Whaley-connell, Adam T.; Sowers, James R.

2011-01-01

108

Regional gene expression of LOX-1, VCAM-1, and ICAM-1 in aorta of HIV-1 transgenic rats  

DEFF Research Database (Denmark)

BACKGROUND: Increased prevalence of atherosclerotic cardiovascular disease in HIV-infected patients has been observed. The cause of this accelerated atherosclerosis is a matter of controversy. As clinical studies are complicated by a multiplicity of risk-factors and a low incidence of hard endpoints, studies in animal models could be attractive alternatives. METHODOLOGY/PRINCIPAL FINDINGS: We evaluated gene expression of lectin-like oxidized-low-density-lipoprotein receptor-1 (LOX-1), vascular cell adhesion molecule-1 (VCAM-1), and intercellular adhesion molecule-1 (ICAM-1) in HIV-1 transgenic (HIV-1Tg) rats; these genes are all thought to play important roles in early atherogenesis. Furthermore, the plasma level of sICAM-1 was measured. We found that gene expressions of LOX-1 and VCAM-1 were higher in the aortic arch of HIV-1Tg rats compared to controls. Also, the level of sICAM-1 was elevated in the HIV-1Tg rats compared to controls, but the ICAM-1 gene expression profile did not show any differences between the groups. CONCLUSIONS/SIGNIFICANCE: HIV-1Tg rats have gene expression patterns indicating endothelial dysfunction and accelerated atherosclerosis in aorta, suggesting that HIV-infection per se may cause atherosclerosis. This transgenic rat model may be a very promising model for further studies of the pathophysiology behind HIV-associated cardiovascular disease.

Hag, Anne Mette Fisker; Kristoffersen, Ulrik Sloth

2009-01-01

109

Rheumatic manifestations of inflammatory bowel disease  

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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...

Rodri?guez-reyna, Tatiana Sofi?a; Marti?nez-reyes, Cynthia; Yamamoto-furusho, Jesu?s Kazu?o

2009-01-01

110

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

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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

111

Temporal sensitivity changes with extended training in a bisection task in a transgenic rat model  

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Full Text Available The present study investigated temporal perception in a Huntington Disease transgenic rat model using a temporal bisection procedure. After initial discrimination training in which animals learned to press one lever after a 2-s tone duration, and the other lever after a 8-s tone duration for food reward, the bisection procedure was implemented in which intermediate durations with no available reinforcement were interspersed with trials with the anchor durations. Bisection tests were repeated in a longitudinal design from 4 to 8 months of age. The results showed that response latencies evolved from a monotonic step-function to an inverted U-shaped function with repeated testing, a precursor of nonresponding on trials with intermediate durations. We inferred that temporal sensitivity and incentive motivation combined to control the transformation of the bisection task from a two-choice task at the outset of testing to a three-choice task with repeated testing. Changes in the structure of the task and/or continued training were accompanied by improvement in temporal sensitivity. In sum, the present data highlight the possible joint roles of temporal and non-temporal factors in the temporal bisection task, and suggested that non-temporal factors may compensate for deficits in temporal processing.

ValerieDoyere

2011-09-01

112

Production and sorting of transgenic, modified human parathyroid hormone in vivo in rat salivary glands.  

Science.gov (United States)

Polarized salivary epithelial cells can sort secretory proteins towards either the basolateral or apical pole. Transgenic human parathyroid hormone (hPTH) exclusively sorts apically in rat submandibular glands. To help understand this specific process we modified the hPTH cDNA sequence and delivered the cDNAs to glands in vivo using adenoviral (Ad) vectors. The Ad vectors encoded: (1) the native form of hPTH (Ad.pre-pro-hPTH1-84), (2) the native sequence, but with the pro-segment deleted (Ad.pre-hPTH1-84), and (3) a sequence containing the pre-segment followed by the first 34 amino acids of hPTH (Ad.pre-hPTH1-34). hPTH production and sorting were studied after two days. All constructs were effectively transcribed in targeted glands. However, the pre-hPTH1-84 modification led to reduced hPTH secretion and production, while no immunoreactive hPTH resulted from pre-hPTH1-34 cDNA infusion. The pre-hPTH1-84 modification had no effect on apical sorting. These in vivo results show that the signal responsible for hPTH's apical sorting does not reside in the pro-segment and that deleting both the pro-segment and the carboxyl-terminal region severely impairs post-translational processing of hPTH. PMID:19944067

Adriaansen, Janik; Zheng, Changyu; Perez, Paola; Baum, Bruce J

2010-01-01

113

Neuronal driven pre-plaque inflammation in a transgenic rat model of Alzheimer's disease.  

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Chronic brain inflammation is associated with Alzheimer's disease (AD) and is classically attributed to amyloid plaque deposition. However, whether the amyloid pathology can trigger early inflammatory processes before plaque deposition remains a matter of debate. To address the possibility that a pre-plaque inflammatory process occurs, we investigated the status of neuronal, astrocytic, and microglial markers in pre- and post-amyloid plaque stages in a novel transgenic rat model of an AD-like amyloid pathology (McGill-R-Thy1-APP). In this model, we found a marked upregulation of several classical inflammatory markers such as COX-2, IL-1?, TNF-?, and fractalkine (CX3CL1) in the cerebral cortex and hippocampus. Interestingly, many of these markers were highly expressed in amyloid beta-burdened neurons. Activated astrocytes and microglia were associated with these A?-burdened neurons. These findings confirm the occurrence of a proinflammatory process preceding amyloid plaque deposition and suggest that A?-burdened neurons play a crucial role in initiating inflammation in AD. PMID:24831823

Hanzel, Cecilia E; Pichet-Binette, Alexa; Pimentel, Luisa S B; Iulita, M Florencia; Allard, Simon; Ducatenzeiler, Adriana; Do Carmo, Sonia; Cuello, A Claudio

2014-10-01

114

Male and female germline specific expression of an EGFP reporter gene in a unique strain of transgenic rats.  

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A rat line was generated in which genomic integration of a ROSA-EGFP transgene resulted in exclusive expression of EGFP in the germ cells of both sexes. EGFP expression was uniform and robust in cleavage stage embryos beginning at the late 2-cell stage and continuing through blastocyst development where expression became restricted to cells of the inner cell mass. Subsequent analysis showed high EGFP expression exclusively in primordial, embryonic, and adult germ cells. This unique expression pattern makes this EGFP marked locus the first molecular marker of the germline lineage in both sexes in mammals. FISH was used to localize the transgene insertion to chromosome 11q11-q12, proximal to Grik1 and near Ncam2. Analysis of the region did not identify known germ cell-specific genes but did identify 19 ESTs or transcribed loci present in testes, ovary, or pre-implantation libraries from mice or rats. To assess the utility of the transgenic line for germ cell transplantation studies, non-selected, freshly isolated seminiferous tubule cells were transferred to the testis of recipient males. The donor cell population colonized the testis at a surprisingly high efficiency within 30 days following transfer. Since EGFP is a vital marker, the colonization process can be followed in vivo and the extent of colonization quantified. The unique germ cell specific expression of EGFP makes this line of transgenic rats an excellent novel tool to study germ cell origin, development, and differentiation, and to assess the plasticity of adult somatic stem cells to become male germ cells. PMID:15993404

Cronkhite, Jennifer T; Norlander, Carola; Furth, Jenny K; Levan, Göran; Garbers, David L; Hammer, Robert E

2005-08-01

115

A 90-day toxicology study of high-amylose transgenic rice grain in Sprague-Dawley rats.  

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A transgenic rice line (TRS) with high amylose level has been developed by antisense RNA inhibition of starch branching enzymes. Compositional analysis of TRS demonstrated that the content of resistant starch (RS) was significantly higher compared to conventional non-transgenic rice. High level of RS is an important raw material in food industry and has various physiological effects for human health. In order to provide the reliable theory basis for field release of TRS rice, we evaluated the potential health effects of long-term consumption of the TRS. The 90-day toxicology feeding experiment was conducted in Sprague-Dawley rats fed with diets containing 70% of either TRS rice flour, its near-isogenic rice flour or the control diet. The clinical performance variables (body weight, body weight gain and food consumption) were measured and pathological responses (hematological parameters and serum chemistry at the midterm and the completion of the experiment, urinalysis profile and serum sex hormone response at the completion of the experiment) were performed. Besides, clinical signs, relative organ weights and microscopic observations were also compared between TRS group and its near-isogenic rice group. The combined data indicates that high-amylose TRS grain is as safe as the conventional non-transgenic rice for rat consumption. PMID:21967780

Zhou, Xing Hua; Dong, Ying; Xiao, Xiang; Wang, Yun; Xu, Yong; Xu, Bin; Shi, Wei Dong; Zhang, Yi; Zhu, Li Jia; Liu, Qiao Quan

2011-12-01

116

Minigenes encoding N-terminal domains of human cardiac myosin light chain-1 improve heart function of transgenic rats.  

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In this study we investigated whether the expression of N-terminal myosin light chain-1 (MLC-1) peptides could improve the intrinsic contractility of the whole heart. We generated transgenic rats (TGR) that overexpressed minigenes encoding the N-terminal 15 amino acids of human atrial MLC-1 (TGR/hALC-1/1-15, lines 7475 and 3966) or human ventricular MLC-1 (TGR/hVLC-1/1-15, lines 6113 and 6114) isoforms in cardiomyocytes. Synthetic N-terminal peptides revealed specific actin binding, with a significantly (Ptransgenic human MLC-1 peptide, but not in a TGR line with undetectable transgene expression levels. The positive inotropic effect of MLC-1 peptides occurred in the absence of a hypertrophic response. Thus, expression of N-terminal domains of MLC-1 represent a valuable tool for the treatment of the failing heart. PMID:16675844

Haase, Hannelore; Dobbernack, Gisela; Tünnemann, Gisela; Karczewski, Peter; Cardoso, Cristina; Petzhold, Daria; Schlegel, Wolfgang-Peter; Lutter, Steffen; Pierschalek, Petra; Behlke, Joachim; Morano, Ingo

2006-05-01

117

Genetic Enhancement of Memory and Long-Term Potentiation but Not CA1 Long-Term Depression in NR2B Transgenic Rats  

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One major theory in learning and memory posits that the NR2B gene is a universal genetic factor that acts as rate-limiting molecule in controlling the optimal NMDA receptor's coincidence-detection property and subsequent learning and memory function across multiple animal species. If so, can memory function be enhanced via transgenic overexpression of NR2B in another species other than the previously reported mouse species? To examine these crucial issues, we generated transgenic rats in whic...

Wang, Deheng; Cui, Zhenzhong; Zeng, Qingwen; Kuang, Hui; Wang, L. Phillip; Tsien, Joe Z.; Cao, Xiaohua

2009-01-01

118

HIV-1 transgene expression in rats induces differential expression of tumor necrosis factor alpha and zinc transporters in the liver and the lung  

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Abstract Background Highly effective antiviral treatment can suppress HIV-1 infection, but the chronic effects of HIV-1-related viral proteins, including gp120 and Tat, on organs such as the lungs can be damaging. HIV-1 transgenic rodent models are useful for studying the systemic effects of these proteins independently of viral infection. We have previously shown that HIV-1 transgene expression (and therefore, HIV-1-related protein expression) in rats decreases alveolar macr...

Joshi Pratibha C; Guidot David M

2011-01-01

119

Roles of Glutathione in Antioxidant Defense, Inflammation, and Neuron Differentiation in the Thalamus of HIV-1 Transgenic Rats.  

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Inflammation and oxidative stress in the brain are major causes of HIV-associated neurocognitive disorders. Previously we have reported high content of glutathione (GSH) in the thalamus of rats with F344 genetic background. In this study, we investigated the changes of GSH metabolism and GSH-dependent antioxidant enzymes in the rat thalamus in response to HIV-1 transgenesis, and their associations with oxidative stress, inflammation, and neuronal development. Male HIV-1 transgenic (HIV-1Tg) rats and wild type F344 rats at 10 months were used in this study, with 5 rats in each group. Parameters measured in this study included: total and oxidized GSH, glutathione peroxidase (GPx), glutathione-S-transferase (GST), gamma-glutamylcysteine synthetase (GCS), gamma-glutamyl transferase (GGT), cysteine/cystine transporters, 4-hydroxynonenal (HNE), interleukin 12 (IL12), neuronal nuclei (NeuN), microtubule-associated protein (MAP2), and glia fibrillary acidic protein (GFAP). The levels of total GSH, oxidized GSH (GSSG) and MAP2 protein, and enzymatic activities of GCS, GPx and GST were significantly higher in HIV-1Tg rats compared with F344 rats, but the ratio of GSSG/GSH, activity of GGT and levels of HNE, NeuN protein and GFAP protein did not change. HIV-1Tg rats showed a lower level of IL12 protein. GSH positively correlated with GCS, GST and MAP2, GSSG/GSH ratio positively correlated with HNE and IL12, the activities of GPx, GST and GCS positively correlated with each other, and negatively correlated with HNE. These findings suggest an important role of the GSH-centered system in reducing oxidative stress and neuroinflammation, and enhancing neuron differentiation in the thalamus of HIV-1Tg rats. PMID:24609977

Pang, Xiaosha; Panee, Jun

2014-06-01

120

The Relationship of Photoreceptor Degeneration to Retinal Vascular Development and Loss in Mutant Rhodopsin Transgenic and RCS Rats  

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The early loss of photoreceptors in some retinal degenerations in mice has been shown to have a profound effect on vascular development of the retina. To better characterize this relationship, we have examined the formation of retinal blood vessels during the first month of life in 8 lines of transgenic rats with different ages of onset and rates of photoreceptor cell loss mediated by the expression of mutant rhodopsin (P23H and S334ter). The number of capillary profiles in the superficial pl...

Pennesi, Mark E.; Nishikawa, Shimpei; Matthes, Michael T.; Yasumura, Douglas; Lavail, Matthew M.

2008-01-01

 
 
 
 
121

Visual Properties of Transgenic Rats Harboring the Channelrhodopsin-2 Gene Regulated by the Thy-1.2 Promoter  

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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 pr...

2009-01-01

122

HIV-1 transgene expression in rats induces differential expression of tumor necrosis factor alpha and zinc transporters in the liver and the lung  

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Full Text Available Abstract Background Highly effective antiviral treatment can suppress HIV-1 infection, but the chronic effects of HIV-1-related viral proteins, including gp120 and Tat, on organs such as the lungs can be damaging. HIV-1 transgenic rodent models are useful for studying the systemic effects of these proteins independently of viral infection. We have previously shown that HIV-1 transgene expression (and therefore, HIV-1-related protein expression in rats decreases alveolar macrophage zinc levels and phagocytic capacity by unknown mechanisms. We hypothesized that HIV-1 transgene expression induces chronic inflammation and zinc sequestration within the liver and thereby decreases zinc bioavailability in the lung. We examined the expression of the pro-inflammatory cytokine, tumor necrosis factor alpha (TNF?, the zinc storage protein, metallothionein (MT1, and the zinc exporter, ZNT1 in the livers and the lungs of wild type and HIV-1 transgenic rats ± dietary zinc supplementation. In addition, we measured zinc levels, the zinc importing protein ZIP1, and the phagocytic capacity in the alveolar macrophages. Results HIV-1 transgene expression increased the liver-specific expression of TNF?, suggesting a chronic inflammatory response within the liver in response to HIV-1-related protein expression. In parallel, HIV-1 transgene expression significantly increased MT1 and ZNT1 expression in the liver as compared to the lung, a pattern that is consistent with zinc sequestration in the liver as occurs during systemic inflammation. Further, HIV-1 transgene expression decreased intracellular zinc levels and increased expression of ZIP1 in the alveolar macrophages, a pattern consistent with zinc deficiency, and decreased their bacterial phagocytic capacity. Interestingly, dietary zinc supplementation in HIV-1 transgenic rats decreased gene expression of TNF?, MT1, and ZNT1 in the liver while simultaneously increasing their expression in the lung. In parallel, zinc supplementation increased alveolar macrophage intracellular zinc levels and bacterial phagocytic capacity in HIV-1 transgenic rats. Conclusion Taken together, these findings suggest that chronic HIV-1-related protein expression causes liver inflammation and zinc sequestration, which in turn limits zinc bioavailability in the lung and thereby impairs alveolar macrophage phagocytic function. Importantly, dietary zinc supplementation decreases liver inflammation and zinc sequestration and restores alveolar macrophage phagocytic function in HIV-1 transgenic rats, a result with potential clinical implications for improving lung health in HIV-1-infected individuals.

Guidot David M

2011-10-01

123

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

DEFF Research Database (Denmark)

Conversion of soluble α-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 α-synuclein conversion and deposition at nigrostriatal dopaminergic synapses in transgenic mice, mainly owing to expression limitations of the α-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 α-synuclein could exert dopaminergic neuropathological effects. We found that the human promoter induced a pan-neuronal expression, matching the rodent α-synuclein expression pattern, however, with prominent C-terminally truncated fragments. Ageing promoted conversion of both full-length and C-terminally truncated α-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

124

[Development of collagen vascular diseases and production of autoantibodies in HTLV-I env-pX transgenic rats].  

Science.gov (United States)

Human T lymphocyte virus type I (HTLV- I) is now known to be associated with a number of diverse clinical disorders, not only adult T cell leukemia but also HTLV- I associated myelopathy/tropical spastic paraparesis, HTLV- I -associated arthropathy, HTLV- I uveitis, and probably Sjögren's syndrome, T cell alveolitis, polymyositis, and infective dermatitis. To investigate virus-host interactions and pathogenetic mechanisms in these diverse disorders, inbred rat, which is susceptible to HTLV- I infection and develops HAM/TSP-like disease by HTLV- I infection, was used as a host of HTLV- I gene transfer model. HTLV- I LTR-env-pX-LTR construct was injected to rat ova, and two lines of the transgenic rat (env-pX rat) were established. Both lines of env-pX rats expressed HTLV- I env and pX genes in various tissues, and developed a wide spectrum of collagen vascular diseases, including chronic destructive arthritis similar to rheumatoid arthritis, necrotizing arteritis mimicking polyarteritis nodosa, myositis, myocarditis, and chronic sialoadenitis and dacryoadenitis resembling Sjögren's syndrome in humans. Thrombosis and thymic atrophy were also observed. These rats showed hyper-gamma globulinemia and a number of autoantibodies, including high titered rheumatoid factors, anti-dsDNA antibodies and anti-cardiolipin antibodies were presented in the serum. Results suggest that the HTLV- I env-pX gene may play a pathogenic role in development of collagen vascular diseases associated with autoimmune phenomenon. The env-pX rat appears to be a suitable animal model for elucidating pathogenetic mechanisms implicated in HTLV- I -induced diseases and also in various collagen vascular diseases of unknown etiology in humans. PMID:8752528

Yamazaki, H

1996-05-01

125

The relationship of photoreceptor degeneration to retinal vascular development and loss in mutant rhodopsin transgenic and RCS rats.  

Science.gov (United States)

The early loss of photoreceptors in some retinal degenerations in mice has been shown to have a profound effect on vascular development of the retina. To better characterize this relationship, we have examined the formation of retinal blood vessels during the first month of life in 8 lines of transgenic rats with different ages of onset and rates of photoreceptor cell loss mediated by the expression of mutant rhodopsin (P23H and S334ter). The number of capillary profiles in the superficial plexus (SP) and deep capillary plexus (DCP) of the retina were quantified in retinal sections taken at postnatal day (P) 8, 10, 12, 15 and 30. In normal wild-type rats, the SP and DCP had mostly established mature, adult patterns by P15, as previously shown. In the transgenic rats, the loss of photoreceptors had relatively little effect on the SP. By contrast, the loss of photoreceptors during vascular development had a major impact on the DCP. In the two lines with early and most rapid photoreceptor loss, S334ter-7 and S334ter-3, where about 90% and 65%, respectively, of the photoreceptors were already lost by P15, the DCP either failed to form (S334ter-7) or the number of capillary profiles was less than 7% of controls (S334ter-3). In lines where almost all photoreceptors were still present at P15 (S334ter-4, S334ter-9, P23H-2 and P23H-3), the number of profiles in the DCP were the same as in wild-type controls at P30. In two lines with an intermediate rate of degeneration (S334ter-5 and P23H-1), where only about 25% of the photoreceptors were lost by P15, there was an intermediate number of vascular profiles in the DCP at P30. Thus, a very close relationship between the number of photoreceptors and vessel profiles in the DCP during its development exists in the transgenic rats, and the loss of photoreceptors results in the failure or inhibition of the DCP to develop. Several mechanisms may explain this relationship including changes in the level of physiological oxygen tension or alteration in the release of angiogenic factors that normally drive vessel development. Analysis of older transgenic retinas up to 1 year of age revealed that (1) vascular profiles are lost from the DCP in essentially all lines once fewer than about 30-33% of photoreceptors remain; (2) in those lines where the DCP essentially did not develop (S334ter-7 and S334ter-3), the effect of photoreceptor absence was permanent, and there was no late vascularization of the DCP; (3) the number of capillary profiles in the SP remained no different from controls in any of the lines, despite long-standing loss of photoreceptors; and (4) neovascularization of the RPE by retinal capillaries occurred with a latency of 60-180 days after the loss of photoreceptors, except in S334ter-7 rats, where neovascularization essentially did not occur. Analysis of RCS rats was carried out for comparison. PMID:18848932

Pennesi, Mark E; Nishikawa, Shimpei; Matthes, Michael T; Yasumura, Douglas; LaVail, Matthew M

2008-12-01

126

Dietary effects of soy isoflavones daidzein and genistein on 7,12-dimethylbenz[a]anthracene-induced mammary mutagenesis and carcinogenesis in ovariectomized Big Blue transgenic rats.  

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The major constituents of isoflavones daidzein (DZ) and genistein (GE) interact with the and estrogen receptors in several tissues including mammary tissues. In this study, we used ovariectomy (OVX) to model menopause and determined the effects of DZ, GE or 17beta-estradiol (E(2)) exposures on chemically induced mutagenesis and carcinogenesis in the mammary glands of female Big Blue transgenic rats. The rats were fed control diet containing the isoflavones and E(2) and treated with a single o...

Manjanatha, Mugimane; Shelton, Sharon; Bishop, Michelle; Lyn-cook, Lascelles; Aidoo, Anane

2006-01-01

127

The direct renin inhibitor aliskiren improves vascular remodelling in transgenic rats harbouring human renin and angiotensinogen genes.  

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In the present study, we tested the hypothesis that chronic treatment with the direct rennin inhibitor aliskiren improves the remodelling of resistance arteries in dTGR (double-transgenic rats). dTGR (5 weeks) were treated with aliskiren (3 mg/kg of body mass per day) or ramipril (1 mg/kg of body mass per day) for 14 days and compared with age-matched vehicle-treated dTGR. BP (blood pressure) was similarly reduced in both aliskiren-treated and ramipril-treated rats compared with control dTGR (167±1 and 169±2 mmHg compared with 197±4 mmHg respectively; Pramipril-treated rats compared with controls (6.3±0.5 and 6.4±0.2% compared with 9.8±0.4% respectively; Pramipril-treated dTGR (Pramipril compared with controls. Only aliskiren induced a 2-fold increase in plasma nitrite, which was significantly greater than that induced by ramipril (Pramipril reduced the M/L ratio of mesenteric arteries and improved oxidative stress in dTGR. However, only aliskiren increased further NO production in the vasculature. Hence, in dTGR, direct renin inhibition induces favourable effects similar to that induced by ACE (angiotensin-converting enzyme) inhibition in improving vascular remodelling through different mechanisms. PMID:23438195

Savoia, Carmine; Arrabito, Emanuele; Parente, Rosa; Sada, Lidia; Madaro, Luca; Nicoletti, Carmine; Zezza, Luigi; Alonzo, Alessandro; Rubattu, Speranza; Michelini, Serena; Muller, Dominik N; Volpe, Massimo

2013-08-01

128

Efficacy of methotrexate in the treatment of a HLA-B27-positive Japanease patient with reactive arthritis.  

Science.gov (United States)

We report a case of reactive arthritis in a 21-year-old man who was successfully treated with methotrexate. In July 2008, the patient experienced arthritis in the left knee 3 days after being diagnosed as having urethritis by the urology clinic. The patient was treated with loxoprofen sodium and fosfomycin calcium at an orthopedic clinic. Antibiotics induced clinical improvement of urethritis, although arthritis became worse. Even after sulphasalazine and corticosteroid were started, polyarthritis remained persistent. Finally, methtrexate was added ; thereafter, polyarthritis and elevated CRP were resolved. HLA-B270502 was positive. Methotrexate could be one of the choices for sulphasalazine-resistant reactive arthritis. PMID:21048390

Nanke, Yuki; Yago, Toru; Kobashigawa, Tsuyoshi; Kotake, Shigeru

2010-01-01

129

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

130

Efficient, Glucose Responsive, and Islet-Specific Transgene Expression by a Modified Rat Insulin Promoter  

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This study was done to improve efficiency and islet specificity of the rat insulin promoter (RIP). Various rat insulin promoter lengths were prepared and tested in vitro to drive luciferase reporter gene expression in INS1-cells, alpha-cells, acinar cells, ductal cells, and fibroblasts. The CMV promoter was used as a positive control. In addition, the DsRed reporter gene was administered in vivo to rat pancreas by ultrasound-targeted microbubble destruction (UTMD). Confocal microscopy was use...

2009-01-01

131

Restoration of visual function in P23H rhodopsin transgenic rats by gene delivery of BiP/Grp78.  

Science.gov (United States)

The P23H mutation within the rhodopsin gene (RHO) causes rhodopsin misfolding, endoplasmic reticulum (ER) stress, and activates the unfolded protein response (UPR), leading to rod photoreceptor degeneration and autosomal dominant retinitis pigmentosa (ADRP). Grp78/BiP is an ER-localized chaperone that is induced by UPR signaling in response to ER stress. We have previously demonstrated that BiP mRNA levels are selectively reduced in animal models of ADRP arising from P23H rhodopsin expression at ages that precede photoreceptor degeneration. We have now overexpressed BiP to test the hypothesis that this chaperone promotes the trafficking of P23H rhodopsin to the cell membrane, reprograms the UPR favoring the survival of photoreceptors, blocks apoptosis, and, ultimately, preserves vision in ADRP rats. In cell culture, increasing levels of BiP had no impact on the localization of P23H rhodopsin. However, BiP overexpression alleviated ER stress by reducing levels of cleaved pATF6 protein, phosphorylated eIF2alpha and the proapoptotic protein CHOP. In P23H rats, photoreceptor levels of cleaved ATF6, pEIF2alpha, CHOP, and caspase-7 were much higher than those of wild-type rats. Subretinal delivery of AAV5 expressing BiP to transgenic rats led to reduction in CHOP and photoreceptor apoptosis and to a sustained increase in electroretinogram amplitudes. We detected complexes between BiP, caspase-12, and the BH3-only protein BiK that may contribute to the antiapoptotic activity of BiP. Thus, the preservation of photoreceptor function resulting from elevated levels of BiP is due to suppression of apoptosis rather than to a promotion of rhodopsin folding. PMID:20231467

Gorbatyuk, Marina S; Knox, Tessa; LaVail, Matthew M; Gorbatyuk, Oleg S; Noorwez, Syed M; Hauswirth, William W; Lin, Jonathan H; Muzyczka, Nicholas; Lewin, Alfred S

2010-03-30

132

Restoration of visual function in P23H rhodopsin transgenic rats by gene delivery of BiP/Grp78  

Science.gov (United States)

The P23H mutation within the rhodopsin gene (RHO) causes rhodopsin misfolding, endoplasmic reticulum (ER) stress, and activates the unfolded protein response (UPR), leading to rod photoreceptor degeneration and autosomal dominant retinitis pigmentosa (ADRP). Grp78/BiP is an ER-localized chaperone that is induced by UPR signaling in response to ER stress. We have previously demonstrated that BiP mRNA levels are selectively reduced in animal models of ADRP arising from P23H rhodopsin expression at ages that precede photoreceptor degeneration. We have now overexpressed BiP to test the hypothesis that this chaperone promotes the trafficking of P23H rhodopsin to the cell membrane, reprograms the UPR favoring the survival of photoreceptors, blocks apoptosis, and, ultimately, preserves vision in ADRP rats. In cell culture, increasing levels of BiP had no impact on the localization of P23H rhodopsin. However, BiP overexpression alleviated ER stress by reducing levels of cleaved pATF6 protein, phosphorylated eIF2? and the proapoptotic protein CHOP. In P23H rats, photoreceptor levels of cleaved ATF6, pEIF2?, CHOP, and caspase-7 were much higher than those of wild-type rats. Subretinal delivery of AAV5 expressing BiP to transgenic rats led to reduction in CHOP and photoreceptor apoptosis and to a sustained increase in electroretinogram amplitudes. We detected complexes between BiP, caspase-12, and the BH3-only protein BiK that may contribute to the antiapoptotic activity of BiP. Thus, the preservation of photoreceptor function resulting from elevated levels of BiP is due to suppression of apoptosis rather than to a promotion of rhodopsin folding.

Gorbatyuk, Marina. S.; Knox, Tessa; LaVail, Matthew M.; Gorbatyuk, Oleg S.; Noorwez, Syed M.; Hauswirth, William W.; Lin, Jonathan H.; Muzyczka, Nicholas; Lewin, Alfred S.

2010-01-01

133

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

2006-09-30

134

Potassium 2-(1-hydroxypentyl)-benzoate promotes long-term potentiation in A?1-42-injected rats and APP/PS1 transgenic mice.  

Science.gov (United States)

Aim:Potassium 2-(1-hydroxypentyl)-benzoate (dl-PHPB) is a new drug candidate for ischemic stroke. The aim of this study was to investigate the effects of dl-PHPB on memory deficits and long-term potentiation (LTP) impairment in animal models of Alzheimer's disease.Methods:The expression of NMDA receptor subunits GluN1 and GluN2B in the hippocampus and cortex of APP/PS1 transgenic mice were detected using Western blot analysis. Memory deficits of the mice were evaluated with the passive avoidance test. LTP impairment was studied in the dentate region of A?1-42-injected rats and APP/PS1 transgenic mice.Results:APP/PS1 transgenic mice showed significantly lower levels of GluN1 and p-GluN2B in hippocampus, and chronic administration of dl-PHPB (100 mg·kg(-1)·d(-1), po) reversed the downregulation of p-GluN2B, but did not change GluN1 level in the hippocampus. Furthermore, chronic administration of dl-PHPB reversed the memory deficits in APP/PS1 transgenic mice. In the dentate region of normal rats, injection of dl-PHPB (100 ?mol/L, icv) did not change the basal synaptic transmission, but significantly enhanced the high-frequency stimulation (HFS)-induced LTP, which was completely prevented by pre-injection of APV (150 ?mol/L, icv). Chronic administration of dl-PHPB (100 mg·kg(-1)·d(-1), po) reversed LTP impairment in A?1-42-injected normal rats and APP/PS1 transgenic mice.Conclusion:Chronic administration of dl-PHPB improves learning and memory and promotes LTP in the animal models of Alzheimer's disease, possibly via increasing p-GluN2B expression in the hippocampus. PMID:24858312

Li, Ping-Ping; Wang, Wei-Ping; Liu, Zhi-Hui; Xu, Shao-Feng; Lu, Wen-Wen; Wang, Ling; Wang, Xiao-Liang

2014-07-01

135

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

Science.gov (United States)

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

136

Time and time again: temporal processing demands implicate perceptual and gating deficits in the HIV-1 transgenic rat.  

Science.gov (United States)

HIV-1-associated neurocognitive disorders (HAND) afflict up to 50 % of HIV-1+ individuals, despite the effectiveness of combination antiretroviral therapy (CART) in reducing the prevalence of more severe neurocognitive impairment. Alterations in brainstem auditory evoked potentials (BAEP), a measure of temporal processing, are one of the earliest neurological abnormalities of HIV-1-positive individuals. Prepulse inhibition (PPI) of the auditory startle response (ASR), a measure of sensorimotor gating, was studied in HIV-1 transgenic (Tg) rats, which express 7 of the 9 HIV-1 genes. Ovariectomized female Fischer HIV-1 Tg and control rats (ns?=?41-42) were tested for PPI at three test periods, with at least 2 months separating each test period, using auditory and visual prepulses, an auditory startle stimulus, and interstimulus intervals (ISI) ranging from 0 to 4000 msec. Auditory and visual prepulse trial blocks were presented in counterbalanced order. For both auditory and visual prepulses, HIV-1 Tg animals exhibited a flatter ISI function, which did not sharpen with age, as it did in controls. Over time, auditory prepulses precipitated a temporal shift in peak inhibition in HIV-1 Tg animals relative to controls, whereas with visual prepulses, both groups displayed peak inhibition at the 40 msec ISI. A lack of perceptual sharpening with age and a relative insensitivity to the temporal dimension of sensorimotor gating are evident in the HIV-1 Tg rat prior to clinical signs of wasting. Deficits in sensorimotor gating may not only provide an early subtle diagnostic marker of HAND, but may also afford a key target for development of potential therapeutics. PMID:23690140

Moran, Landhing M; Booze, Rosemarie M; Mactutus, Charles F

2013-09-01

137

Transgenic Expression of Aflatoxin Aldehyde Reductase (AKR7A1) Modulates Aflatoxin B1 Metabolism but not Hepatic Carcinogenesis in the Rat  

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In both experimental animals and humans, aflatoxin B1 (AFB1) is a potent hepatic toxin and carcinogen against which a variety of antioxidants and experimental or therapeutic drugs (e.g., oltipraz, related dithiolethiones, and various triterpenoids) protect from both acute toxicity and carcinogenesis. These agents induce several hepatic glutathione S-transferases (GST) as well as aldo-keto reductases (AKR) which are thought to contribute to protection. Studies were undertaken in transgenic rat...

Roebuck, Bill D.; Johnson, Denise N.; Sutter, Carrie Hayes; Egner, Patricia A.; Scholl, Peter F.; Friesen, Marlin D.; Baumgartner, Karen J.; Ware, Nicholas M.; Bodreddigari, Sridevi; Groopman, John D.; Kensler, Thomas W.; Sutter, Thomas R.

2009-01-01

138

Milk Products Containing Bioactive Tripeptides Have an Antihypertensive Effect in Double Transgenic Rats (dTGR) Harbouring Human Renin and Human Angiotensinogen Genes  

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Tripeptides isoleucyl-prolyl-proline (IPP) and valyl-prolyl-proline (VPP) act as ACE inhibitors in vitro. Double transgenic rats (dTGR) harbouring human renin and human angiotensinogen genes develop malignant hypertension due to increased angiotensin II formation. The present study was aimed to evaluate possible antihypertensive effect of IPP and VPP in this severe model. Four-week-old dTGR were randomized in three groups to receive: (1) water (control), (2) fermented milk containing IPP and ...

2010-01-01

139

Milk products containing bioactive tripeptides have an antihypertensive effect in double transgenic rats (dTGR) harbouring human renin and human angiotensinogen genes  

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Tripeptides isoleucyl-prolyl-proline (IPP) and valyl-prolyl-proline (VPP) act as ACE inhibitors in vitro. Double transgenic rats (dTGR) harbouring human renin and human angiotensinogen genes develop malignant hypertension due to increased angiotensin II formation. The present study was aimed to evaluate possible antihypertensive effect of IPP and VPP in this severe model. Four-week-old dTGR were randomized in three groups to receive: (1) water (control), (2) fermented milk containing IPP and ...

2010-01-01

140

Decreased cardiac SERCA2 expression, SR Ca uptake, and contractile function in hypothyroidism are attenuated in SERCA2 overexpressing transgenic rats  

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The sarco/endoplasmic reticulum (SR) Ca2+-ATPase SERCA2a has a key role in controlling cardiac contraction and relaxation. In hypothyroidism, decreased expression of the thyroid hormone (TH)-responsive SERCA2 gene contributes to slowed SR Ca2+ reuptake and relaxation. We investigated whether cardiac expression of a TH-insensitive SERCA2a cDNA minigene can rescue SR Ca2+ handling and contractile function in female SERCA2a-transgenic rats (TG) with experimental hypothyroidism. Wild-type rats (W...

Vetter, Roland; Rehfeld, Uwe; Reissfelder, Christoph; Fechner, Henry; Seppet, Enn; Kreutz, Reinhold

2011-01-01

 
 
 
 
141

Ribozyme rescue of photoreceptor cells in P23H transgenic rats: long-term survival and late-stage therapy.  

Science.gov (United States)

Ribozyme-directed cleavage of mutant mRNAs appears to be a potentially effective therapeutic measure for dominantly inherited diseases. We previously demonstrated that two ribozymes targeted to the P23H mutation in rhodopsin slow photoreceptor degeneration in transgenic rats for up to 3 months of age when injected before significant degeneration at postnatal day (P) 15. We now have explored whether ribozyme rescue persists at older ages, and whether ribozymes are effective when injected later in the degeneration after significant photoreceptor cell loss. Recombinant adeno-associated virus (rAAV) vectors incorporating a proximal bovine rod opsin promoter were used to transfer either hairpin or hammerhead ribozyme genes to photoreceptors. For the study of long-term survival, rAAV was administered by subretinal injection at P15, and the rats were allowed to live up to 8 months of age. For the study of late-stage gene transfer, rAAV was administered at P30 or P45, when 40-45% of the photoreceptors already had degenerated. Eyes were examined functionally by the electroretinogram and structurally by morphometric analysis. When injected at P15, expression of either ribozyme markedly slowed the rate of photoreceptor degeneration for at least 8 months and resulted in significantly greater electroretinogram amplitudes at least up to P180. When injected at P30 or P45, virtually the same number of photoreceptors survived at P130 as when injected at P15. Ribozyme rescue appears to be a potentially effective, long-term therapy for autosomal dominant retinal degeneration and is highly effective even when the gene transfer is done after significant photoreceptor cell loss. PMID:11005848

LaVail, M M; Yasumura, D; Matthes, M T; Drenser, K A; Flannery, J G; Lewin, A S; Hauswirth, W W

2000-10-10

142

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

Science.gov (United States)

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. PMID:23462119

Heijnen, Bart F J; Pelkmans, Leonie P J; Danser, A H Jan; Garrelds, Ingrid M; Mullins, John J; De Mey, Jo G R; Struijker-Boudier, Harry A J; Janssen, Ben J A

2014-03-01

143

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

Science.gov (United States)

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

144

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

145

TRANSGENE-MEDIATED EXPRESSION OF TUMOR NECROSIS FACTOR SOLUBLE RECEPTOR ATTENUATES MORPHINE TOLERANCE IN RATS  

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Opiate/narcotic analgesics are the most effective treatments for chronic severe pain, but their clinical utility is often hampered by the development of analgesic tolerance. Recent evidence suggests chronic morphine may activate glial cells to release proinflammatory cytokines. In this study, we used herpes simplex virus (HSV) vectors-based gene transfer to dorsal root ganglion to produce a local release of p55 TNF soluble receptor in the spinal cord in rats with morphine tolerance. Subcutane...

Sun, Jingxia; Liu, Shue; Mata, Marina; Fink, David J.; Hao, Shuanglin

2012-01-01

146

Fumaric Acid Esters Can Block Pro-Inflammatory Actions of Human CRP and Ameliorate Metabolic Disturbances in Transgenic Spontaneously Hypertensive Rats  

Science.gov (United States)

Inflammation and oxidative stress have been implicated in the pathogenesis of metabolic disturbances. Esters of fumaric acid, mainly dimethyl fumarate, exhibit immunomodulatory, anti-inflammatory, and anti-oxidative effects. In the current study, we tested the hypothesis that fumaric acid ester (FAE) treatment of an animal model of inflammation and metabolic syndrome, the spontaneously hypertensive rat transgenically expressing human C-reactive protein (SHR-CRP), will ameliorate inflammation, oxidative stress, and metabolic disturbances. We studied the effects of FAE treatment by administering Fumaderm, 10 mg/kg body weight for 4 weeks, to male SHR-CRP. Untreated male SHR-CRP rats were used as controls. All rats were fed a high sucrose diet. Compared to untreated controls, rats treated with FAE showed significantly lower levels of endogenous CRP but not transgenic human CRP, and amelioration of inflammation (reduced levels of serum IL6 and TNF?) and oxidative stress (reduced levels of lipoperoxidation products in liver, heart, kidney, and plasma). FAE treatment was also associated with lower visceral fat weight and less ectopic fat accumulation in liver and muscle, greater levels of lipolysis, and greater incorporation of glucose into adipose tissue lipids. Analysis of gene expression profiles in the liver with Affymetrix arrays revealed that FAE treatment was associated with differential expression of genes in pathways that involve the regulation of inflammation and oxidative stress. These findings suggest potentially important anti-inflammatory, anti-oxidative, and metabolic effects of FAE in a model of inflammation and metabolic disturbances induced by human CRP.

Silhavy, Jan; Zidek, Vaclav; Mlejnek, Petr; Landa, Vladimir; Simakova, Miroslava; Strnad, Hynek; Oliyarnyk, Olena; Skop, Vojtech; Kazdova, Ludmila; Kurtz, Theodore; Pravenec, Michal

2014-01-01

147

Hormone secretion in transgenic rats and electrophysiological activity in their gonadotropin releasing-hormone neurons  

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Expression of GFP in GnRH neurons has allowed for studies of individual GnRH neurons. We have demonstrated previously the preservation of physiological function in male GnRH-GFP mice. In the present study, we confirm using biocytin-filled GFP-positive neurons in the hypothalamic slice preparation that GFP-expressing somata, axons, and dendrites in hypothalamic slices from GnRH-GFP rats are GnRH1 peptide positive. Second, we used repetitive sampling to study hormone secretion from GnRH-GFP tra...

Gay, Vernon L.; Hemond, Peter J.; Schmidt, Deena; O Boyle, Michael P.; Hemond, Zoe; Best, Janet; O Farrell, Laura; Suter, Kelly J.

2012-01-01

148

Augmented rod bipolar cell function in partial receptor loss: an ERG study in P23H rhodopsin transgenic and aging normal rats.  

Science.gov (United States)

Physiological consequences of early stages of photoreceptor degeneration were examined in heterozygous P23H rhodopsin transgenic (Tg) and in aging normal Sprague-Dawley rats. Rod photoreceptor and rod bipolar (RB) cell function were estimated with maximum value and sensitivity parameters of P3 and P2 components of the electroretinogram. In both Tg and aging normal rats, the age-related rate of decline of P3 amplitude was steeper than that of the P2 amplitude. Tg rats showed greater than normal sensitivity of the rods. A new model of distal RB pathway connectivity suggested photoreceptor loss could not be the sole cause of physiological abnormalities; there was an additional increase of post-receptoral sensitivity. We propose that changes at rod-RB synapses compensate for the partial loss of rod photoreceptors in senescence and in early stages of retinal degeneration. PMID:11587727

Aleman, T S; LaVail, M M; Montemayor, R; Ying, G; Maguire, M M; Laties, A M; Jacobson, S G; Cideciyan, A V

2001-09-01

149

Activation of transgenic estrogen receptor-beta by selected phytoestrogens in a stably transduced rat serotonergic cell line.  

Science.gov (United States)

Many flavonoids, a major group of phenolic plant-derived secondary metabolites, are known to possess estrogen-like bioactivities. However, little is known about their estrogenic properties in the central nervous system due to the lack of suitable cellular models expressing sufficient amounts of functional estrogen receptor beta (ERbeta). To overcome this deficit, we have created a cellular model, which is serotonergic in origin, to study properties of estrogenic substances by stably transducing RN46A-B14 cells derived from raphe nuclei region of the rat brain with a lentiviral vector encoding a human ERbeta. We clearly showed that the transgenic human ERbeta is a spontaneously expressed and a functional receptor. We have further assessed the estrogenicity of three different isoflavones and four different naringenin-type flavanones in this cell line utilizing a luciferase reporter gene assay. Genistein (GEN), Daidzein (DAI), Equol (EQ), Naringenin (NAR) and 8-prenylnaringenin (8-PN) showed strong estrogenic activity in a concentration-dependent manner as compared to 7-(O-prenyl)naringenin-4'-acetate (7-O-PN) which was only slightly estrogenic and 6-(1,1-dimethylallyl)naringenin (6-DMAN) that neither showed estrogenic nor anti-estrogenic activity in our model. All observed effects could be antagonized by the anti-estrogen fulvestrant. Moreover, co-treatment of cells with 17beta-estradiol (E2) and either GEN or DAI showed a slight additive effect as compared to EQ. On the other hand, 8-PN in addition to 7-O-PN, but not NAR and 6-DMAN, were able to slightly antagonize the responses triggered by E2. Our newly established cellular model may prove to be a useful tool in explicating basic physiological properties of ERbeta in the brain and may help unravel molecular and cellular mechanisms involved in serotonergic mood regulation by estrogen or potential plant-derived secondary metabolites. PMID:20433925

Amer, Dena A M; Kretzschmar, Georg; Müller, Nicole; Stanke, Nicole; Lindemann, Dirk; Vollmer, Günter

2010-06-01

150

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

151

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

International Nuclear Information System (INIS)

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 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.

2009-04-15

152

(1)H NMR based metabolomics of CSF and blood serum: a metabolic profile for a transgenic rat model of Huntington disease.  

Science.gov (United States)

Huntington disease (HD) is a hereditary brain disease. Although the causative gene has been found, the exact mechanisms of the pathogenesis are still unknown. Recent investigations point to metabolic and energetic dysfunctions in HD neurons. Both univariate and multivariate analyses were used to compare proton nuclear magnetic resonance spectra of serum and cerebrospinal fluid (CSF) taken from presymptomatic HD transgenic rats and their wild-type littermates. N-acetylaspartate (NAA), was found to be significantly decreased in the serum of HD rats compared to wild-type littermates. Moreover, in the serum their levels of glutamine, succinic acid, glucose and lactate are significantly increased as well. An increased concentration of lactate and glucose is also found in CSF. There is a 1:1 stoichiometry coupling glucose utilization and glutamate cycling. The observed increase in the glutamine concentration, which indicates a shutdown in the neuronal-glial glutamate-glutamine cycling, results therefore in an increased glucose concentration. The elevated succinic acid concentration might be due to an inhibition of succinate dehydrogenase, an enzyme linked to the mitochondrial respiratory chain and TCA cycle. Moreover, reduced levels of NAA may reflect an impairment of mitochondrial energy production. In addition, the observed difference in lactate supports a deficiency of oxidative energy metabolism in rats transgenic for HD as well. The observed metabolic alterations seem to be more profound in serum than in CSF in presymptomatic rats. All findings suggest that even in presymptomatic rats, a defect in energy metabolism is already apparent. These results support the hypothesis of mitochondrial energy dysfunction in HD. PMID:21867751

Verwaest, Kim A; Vu, Trung N; Laukens, Kris; Clemens, Laura E; Nguyen, Huu P; Van Gasse, Bjorn; Martins, José C; Van Der Linden, Annemie; Dommisse, Roger

2011-11-01

153

Safety assessment of transgenic Bacillus thuringiensis rice T1c-19 in Sprague-Dawley rats from metabonomics and bacterial profile perspectives.  

Science.gov (United States)

Bacillus thuringiensis rice is facing commercialization as the main food source in the near future. The unintended effects of genetically modified (GM) organisms are the most important barriers to their promotion. We aimed to establish a new in vivo evaluation model for genetically modified foods by using metabonomics and bacterial profile approaches. T1c-19 rice flour or its transgenic parent MH63 was used at 70% wt/wt to produce diets that were fed to rats for ? 90 days. Urine metabolite changes were detected using (1)H NMR. Denaturing gradient gel electrophoresis and real-time polymerase chain reaction (RT-PCR) were used to detect the bacterial profiles between the two groups. The metabonomics was analyzed for metabolite changes in rat urine, when compared with the non-GM rice group, where rats were fed a GM rice diet. Several metabolites correlated with rat age and sex but not with GM rice diet. Significant biological differences were not identified between the GM rice diet and the non-GM rice diet. The bacteria related to rat urine metabolites were also discussed. The results from metabonomics and bacterial profile analyses were comparable with the results attained using the traditional method. Because metabonomics and bacterial profiling offer noninvasive, dynamic approaches for monitoring food safety, they provide a novel process for assessing the safety of GM foods. PMID:22215564

Cao, Sishuo; He, Xiaoyun; Xu, Wentao; Luo, YunBo; Yuan, Yanfang; Liu, Pengfei; Cao, Bo; Shi, Hui; Huang, Kunlun

2012-03-01

154

Left ventricular mechanical and energetic changes in long-term isoproterenol-induced hypertrophied hearts of SERCA2a transgenic rats.  

Science.gov (United States)

Overexpression of cardiac sarcoplasmic reticulum Ca(2+)-ATPase (SERCA2a) has been suggested as a strategic intervention for cardiac failure. However, its benefit in wild-type (WT) rats with normal SERCA2a levels seems to be small. To investigate whether it would be beneficial in a cardiac failure model with down-regulated SERCA2a levels, we made a cardiac hypertrophy model using isoproterenol infusion (1.2mgkg(-1)day(-1) for 1 or 4weeks; TG-ISO1w and TG-ISO4w, respectively) in SERCA2a transgenic (TG) rats and compared these rats with littermate WT rats that underwent the same treatments (WT-ISO1w and WT-ISO4w). We analyzed the left ventricular (LV) mechanoenergetics in the excised heart using our original cross-circulation system. The downward shift of curvilinear LV end-systolic pressure-volume relations (ESPVRs) observed in WT-ISO4w rats was abolished in TG-ISO4w rats. The slope and VO2 intercept of the VO2 (myocardial oxygen consumption per beat)-PVA (systolic pressure-volume area: total mechanical energy per beat) linear relation did not differ in any of the groups. The most important finding was a significantly smaller O2 cost of LV contractility in the TG-ISO4w group, which means that less O2 is needed to exert the same LV contractility, compared with the other groups. The increased ratio of SERCA2a/phospholamban returned to the level of the WT-control group only in the TG-ISO4w group. Longer-term up-regulation of mitochondrial transcription factor A for genes of mitochondrial enzymes producing ATP was observed in TG rats. In conclusion, longer-term overexpression of SERCA2a will be beneficial in the present cardiac failure model with down-regulated SERCA2a levels. PMID:23458361

Mitsuyama, Shinichi; Takeshita, Daisuke; Obata, Koji; Zhang, Guo-Xing; Takaki, Miyako

2013-06-01

155

ERK activation induced by selenium treatment significantly downregulates beta/gamma-secretase activity and Tau phosphorylation in the transgenic rat overexpressing human selenoprotein M.  

Science.gov (United States)

Selenium reportedly contribute to the modulation process of protein phosphorylation to regulate various cellular functions including growth, differentiation, proliferation and development. The aim of this study was to investigate whether selenium and Selenoprotein M (SelM) affects the mechanism of Alzheimer's disease. To achieve this, we determined the change of the MAPK pathway, secretase activity, and Tau phosphorylation in the transgenic rat overexpressing human selenoprotein M. Based on these results, we concluded that, i) CMV/GFP-hSelM Tg rats showed a high activity level of antioxidant enzyme in the brain tissues, ii) in response to selenium treatment, the ERK signaling pathway was significantly increased in Tg rats, but did not change in wild-type rats, iii) the activation of the ERK pathway by selenium treatment and SelM overexpression induced the inhibition of the alpha/gamma-secretase activity related to the protection of Abeta-42 production, iv) the activation of the ERK pathway by selenium treatment and SelM overexpression inhibited the phosphorylation in several sites of Tau protein. Therefore, these results provide strong evidence that selenium treatment and SelM activate the ERK pathway to attenuate alpha/gamma-secretase-mediated proteolysis and Tau phosphorylation to protect brain function. PMID:19513540

Yim, Su Y; Chae, Kab R; Shim, Sun B; Hong, Jin T; Park, Jung Y; Lee, Chung Y; Son, Hong J; Sheen, Yhun Y; Hwang, Dae Y

2009-07-01

156

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

157

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.

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

158

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...

2006-01-01

159

Free radical trap phenyl-N-tert-butylnitrone protects against light damage but does not rescue P23H and S334ter rhodopsin transgenic rats from inherited retinal degeneration.  

Science.gov (United States)

Phenyl-N-tert-butylnitrone (PBN) protects rat retinas against light damage. Because the degenerative process involved in light damage and inherited retinal degeneration both lead to a common final cell death, apoptosis, we used transgenic rats with a P23H or S334ter rhodopsin mutation to test the effects of PBN on retinal degeneration and light damage and the susceptibility of the transgenic rats to light damage. In the first study, 3-week-old mutant and wild-type rats were given no drug, 0.25% PBN in drinking water, or 0.25% PBN in drinking water plus three daily intraperitoneal injections of PBN (100 mg/kg, i.p., every 8 hr). Electroretinograms were recorded at postnatal day 49, after which the rats were killed for morphometric analysis. There was no photoreceptor rescue by PBN in P23H or S334ter rats, as evidenced by equivalent loss of function and photoreceptor cells in the three treatment groups. In the second study, P23H, S334ter, and wild-type rats were exposed for 24 hr to 2700 lux light. The rats were untreated or treated with PBN (50 mg/kg per injection, every 6 hr, starting before exposure). ERGs were recorded before and 1 d after exposure. Animals were killed 6 d later for morphometric analysis. PBN protected wild-type and P23H but not S334ter retinas from light damage. S334ter retinas were relatively less susceptible to light damage than P23H and wild-type rats. The results suggest that the initiating event(s) that causes photoreceptor cell death in the mutated rats is different from that which occurs in light damage, although both ultimately undergo an apoptotic cell death. PMID:12853423

Ranchon, Isabelle; LaVail, Matthew M; Kotake, Yashige; Anderson, Robert E

2003-07-01

160

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

 
 
 
 
161

HLA-B73: An atypical HLA-B molecule carrying a Bw6-epitope motif variant and a B pocket identical to HLA-B27  

Energy Technology Data Exchange (ETDEWEB)

HLA-B73, first described by Mayr and Kirnbauer (1981), is a poorly characterized allospecificity, serologically related to the B7-CREG. We polymerase chain reaction-amplified, cloned and sequenced the HLA-B alleles of the B-LCL LE023, established from a Spanish Caucasoid individual expressing HLA-B73. 5 refs., 2 figs.

Vilches, C.; Pablo, R. de; Herrero, M.J.; Moreno, M.E.; Kreisler, M. [Hospital Puerta de Hierro, Madrid (Spain)

1994-12-31

162

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

163

Interaction pattern of Arg 62 in the A-pocket of differentially disease-associated HLA-B27 subtypes suggests distinct TCR binding modes.  

Science.gov (United States)

The single amino acid replacement Asp116His distinguishes the two subtypes HLA-B*2705 and HLA-B*2709 which are, respectively, associated and non-associated with Ankylosing Spondylitis, an autoimmune chronic inflammatory disease. The reason for this differential association is so far poorly understood and might be related to subtype-specific HLA:peptide conformations as well as to subtype/peptide-dependent dynamical properties on the nanoscale. Here, we combine functional experiments with extensive molecular dynamics simulations to investigate the molecular dynamics and function of the conserved Arg62 of the ?1-helix for both B27 subtypes in complex with the self-peptides pVIPR (RRKWRRWHL) and TIS (RRLPIFSRL), and the viral peptides pLMP2 (RRRWRRLTV) and NPflu (SRYWAIRTR). Simulations of HLA:peptide systems suggest that peptide-stabilizing interactions of the Arg62 residue observed in crystal structures are metastable for both B27 subtypes under physiological conditions, rendering this arginine solvent-exposed and, probably, a key residue for TCR interaction more than peptide-binding. This view is supported by functional experiments with conservative (R62K) and non-conservative (R62A) B*2705 and B*2709 mutants that showed an overall reduction in their capability to present peptides to CD8+ T cells. Moreover, major subtype-dependent differences in the peptide recognition suggest distinct TCR binding modes for the B*2705 versus the B*2709 subtype. PMID:22403718

Nurzia, Elisa; Narzi, Daniele; Cauli, Alberto; Mathieu, Alessandro; Tedeschi, Valentina; Caristi, Silvana; Sorrentino, Rosa; Böckmann, Rainer A; Fiorillo, Maria Teresa

2012-01-01

164

Multiple, Non-conserved, Internal Viral Ligands Naturally Presented by HLA-B27 in Human Respiratory Syncytial Virus-infected Cells*  

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Cytotoxic T lymphocyte (CTL)-mediated death of virus-infected cells requires prior recognition of short viral peptide antigens that are presented by human leukocyte antigen (HLA) class I molecules on the surface of infected cells. The CTL response is critical for the clearance of human respiratory syncytial virus (HRSV) infection. Using mass spectrometry analysis of complex HLA-bound peptide pools isolated from large amounts of HRSV-infected cells, we identified nine naturally processed HLA-B...

Infantes, Susana; Lorente, Elena; Barnea, Eilon; Beer, Ilan; Cragnolini, Juan Jose?; Garci?a, Ruth; Lasala, Fa?tima; Jime?nez, Mercedes; Admon, Arie; Lo?pez, Daniel

2010-01-01

165

Development of oral epithelial cell line ROE2 with differentiation potential from transgenic rats harboring temperature-sensitive simian virus40 large T-antigen gene.  

Science.gov (United States)

We have developed an immortalized oral epithelial cell line, ROE2, from fetal transgenic rats harboring temperature-sensitive simian virus 40 large T-antigen gene. The cells grew continuously at either a permissive temperature of 33°C or an intermediate temperature of 37°C. At the nonpermissive temperature of 39°C, on the other hand, growth decreased significantly, and the Sub-G1 phase of the cell cycle increased, indicating that the cells undergo apoptosis at a nonpermissive temperature. Histological and immunocytochemical analyses revealed that ROE2 cells at 37°C had a stratified epithelial-like morphology and expressed cytokeratins Krt4 and Krt13, marker proteins for oral nonkeratinized epithelial cells. Global-scale comprehensive microarray analysis, coupled with bioinformatics tools, demonstrated a significant gene network that was obtained from the upregulated genes. The gene network contained 16 genes, including Cdkn1a, Fos, Krt13, and Prdm1, and was associated mainly with the biological process of skin development in the category of biological functions, organ development. These four genes were validated by quantitative real-time polymerase chain reaction, and the results were nearly consistent with the microarray data. It is therefore anticipated that this cell line will be useful as an in vitro model for studies such as physiological functions, as well as for gene expression in oral epithelial cells. PMID:24521861

Tabuchi, Yoshiaki; Wada, Shigehito; Ikegame, Mika; Kariya, Ayako; Furusawa, Yukihiro; Hoshi, Nobuhiko; Yunoki, Tatsuya; Suzuki, Nobuo; Takasaki, Ichiro; Kondo, Takashi; Suzuki, Yoshihisa

2014-01-01

166

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

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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...

Lorber, Barbara; Tassoni, Alessia; Bull, Natalie D.; Moschos, Marilita M.; Martin, Keith R.

2012-01-01

167

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.

TomasPetrasek

2014-03-01

168

Defining Peripheral Nervous System Dysfunction in the SOD-1G93A Transgenic Rat Model of Amyotrophic Lateral Sclerosis.  

Science.gov (United States)

Growing evidence indicates that alterations within the peripheral nervous system (PNS) are involved at an early stage in the amyotrophic lateral sclerosis (ALS) pathogenetic cascade. In this study, magnetic resonance imaging (MRI), neurophysiologic analyses, and histologic analyses were used to monitor the extent of PNS damage in the hSOD-1 ALS rat model. The imaging signature of the disease was defined using in vivo MRI of the sciatic nerve. Initial abnormalities were detected in the nerves by an increase in T2 relaxation time before the onset of clinical disease; diffusion MRI showed a progressive increase in mean and radial diffusivity and reduction of fractional anisotropy at advanced stages of disease. Histologic analysis demonstrated early impairment of the blood-nerve barrier followed by acute axonal degeneration associated with endoneurial edema and macrophage response in motor nerve compartments. Progressive axonal degeneration and motor nerve fiber loss correlated with MRI and neurophysiologic changes. These functional and morphologic investigations of the PNS might be applied in following disease progression in preclinical therapeutic studies. This study establishes the PNS signature in this rat ALS model (shedding new light into pathogenesis) and provides a rationale for translating into future systematic MRI studies of PNS involvement in patients with ALS. PMID:24918640

Riva, Nilo; Chaabane, Linda; Peviani, Marco; Ungaro, Daniela; Domi, Teuta; Dina, Giorgia; Bianchi, Francesca; Spano, Giorgia; Cerri, Federica; Podini, Paola; Corbo, Massimo; Carro, Ubaldo Del; Comi, Giancarlo; Bendotti, Caterina; Quattrini, Angelo

2014-07-01

169

Overexpression of human selenoprotein M differentially regulates the concentrations of antioxidants and H2O2, the activity of antioxidant enzymes, and the composition of white blood cells in a transgenic rat.  

Science.gov (United States)

Selenoprotein is associated with a variety of serious diseases, including infectious diseases, neurodegenerative disorders, cancer and cardiovascular disease. The aim of this study was to produce a new transgenic (Tg) rat expressing human selenoprotein M (SelM) in order to examine the protective function of the antioxidant status in vivo. To achieve this, a new lineage of Tg rats was produced by the microinjection of pCMV/GFP-hSelM constructs into a fertilized rat egg. Several conclusions can be drawn based on the results of the present study. The human SelM gene was successfully expressed at both the transcription and protein levels in the CMV/GFP-hSelM Tg rats. This Tg rat showed a different enzyme activity for the antioxidant protein in the various tissues examined. In response to the 2,2'-azobiz(2-amidinopropane) dihydrochloride (AAPH) injection, the Tg rats showed a lower level of antioxidant and H2O2 concentration as the activity of the antioxidant enzyme was maintained at a higher level in the Tg rats than in the non-Tg rats. Also, the neutrophil-to-lymphocyte ratio was significantly increased in this Tg rat, even though the level of corticosterone remained unchanged in both genotypes. Thus, the results of this study demonstrated that the CMV/GFP-hSelM Tg rat can serve as an animal model for the maintenance of a high level of antioxidant status and can be used to study the biological function of selenoprotein in infectious diseases, cardiovascular disease and cancer. PMID:18204783

Hwang, Dae Youn; Sin, Ji Soon; Kim, Min Sun; Yim, Su Youn; Kim, Yong Kyu; Kim, Chuel Kyu; Kim, Byoung Guk; Shim, Sun Bo; Jee, Seung Wan; Lee, Su Hae; Bae, Chang Joon; Lee, Byoung Chun; Jang, Mee Kyung; Cho, Jung Sik; Chae, Kab Ryong

2008-02-01

170

Epithelial cell differentiation in normal and transgenic mouse intestinal isografts  

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Transgenes consisting of segments of the rat liver fatty acid-binding protein (L-FABP) gene's 5' non-transcribed domain linked to the human growth hormone (hGH) gene (minus its regulatory elements) have provided useful tools for analyzing the mechanisms that regulate cellular and spatial differentiation of the continuously renewing gut epithelium. We have removed the jejunum from normal and transgenic fetal mice before or coincident with, cytodifferentiation of its epithelium. These segments ...

1991-01-01

171

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...

2003-01-01

172

Transgenic Rescue of SF-1-Null Mice  

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Steroidogenic factor 1 (SF-1, Nr5a1, and Ad4bp) is an orphan nuclear receptor required for adrenal and gonad development and endocrine regulation. To extend our understanding of SF-1 function and the mechanisms controlling its expression, a transgenic rescue strategy was employed to locate important transcriptional control regions and to reveal functional roles of the protein. A rat yeast artificial chromosome containing Ftz-F1, the gene encoding SF-1, was used to generate mice with different...

Karpova, Tatiana; Maran, R. R. M.; Presley, Jeremy; Scherrer, Serge P.; Tejada, Lovella; Heckert, Leslie L.

2005-01-01

173

Chronic immunoneutralization of brain angiotensin-(1-12) lowers blood pressure in transgenic (mRen2)27 hypertensive rats  

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Angiotensin-(1-12) [ANG-(1-12)] is a newly identified peptide detected in a variety of rat tissues, including the brain. To determine whether brain ANG-(1-12) participates in blood pressure regulation, we treated male adult (mRen2)27 hypertensive rats (24–28 wk of age) with Anti-ANG-(1-12) IgG or Preimmune IgG via an intracerebroventricular cannula for 14 days. Immunoneutralization of brain ANG-(1-12) lowered systolic blood pressure (?43 ± 8 mmHg on day 3 and ?26 ± 7 mmHg on day 10 fr...

Isa, Katsunori; Garci?a-espinosa, Maria Antonia; Arnold, Amy C.; Pirro, Nancy T.; Tommasi, Ellen N.; Ganten, Detlev; Chappell, Mark C.; Ferrario, Carlos M.; Diz, Debra I.

2009-01-01

174

Treatment of acute hepatic failure in mice by transplantation of mixed microencapsulation of rat hepatocytes and transgenic human fetal liver stromal cells.  

Science.gov (United States)

Microencapsulation-mediated cell therapy overcomes the immune incompatibility between donor and recipient in transplantation. The aim of this study was to investigate the effects of transplantation of microcapsules containing a mixture of rat hepatocytes and human fetal liver stromal cells (hFLSCs), engineered to produce basic fibroblast growth factor (bFGF), on acute liver failure (ALF) in mice. In vitro experiments showed that different combinations of microencapsulated rat's hepatocytes and stromal cells survive, grow, and function better in three-dimensional conditions. The metabolic activity of rat hepatocytes co-microencapsulated with hFLSCs, particularly when engineered to produce bFGF (FLSCs/bFGF), is significantly higher than that of microcapsules with rat hepatocytes alone. Intraperitoneal transplantation of the encapsulated hepatocytes with FLSCs/bFGF increased the survival rate and improved liver function of an ALF mouse model induced by a 70% partial hepatectomy in BALB/C mice. Moreover, dramatic liver regeneration was observed 2 days after transplantation in the group that received intraperitoneal transplantations of encapsulated hepatocytes with FLSCs/bFGF. Therefore, transplantation of encapsulated hepatocytes and hFLSCs/bFGF may be a promising strategy to treat ALF or related liver diseases. PMID:20121581

Teng, Yue; Wang, Yunfang; Li, Shuangyan; Wang, Wei; Gu, Ruolan; Guo, Xin; Nan, Xue; Ma, Xiaojun; Pei, Xuetao

2010-10-01

175

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

176

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

177

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 ...

2006-01-01

178

Chronic immunoneutralization of brain angiotensin-(1-12) lowers blood pressure in transgenic (mRen2)27 hypertensive rats  

Science.gov (United States)

Angiotensin-(1-12) [ANG-(1-12)] is a newly identified peptide detected in a variety of rat tissues, including the brain. To determine whether brain ANG-(1-12) participates in blood pressure regulation, we treated male adult (mRen2)27 hypertensive rats (24–28 wk of age) with Anti-ANG-(1-12) IgG or Preimmune IgG via an intracerebroventricular cannula for 14 days. Immunoneutralization of brain ANG-(1-12) lowered systolic blood pressure (?43 ± 8 mmHg on day 3 and ?26 ± 7 mmHg on day 10 from baseline, P < 0.05). Water intake was lower on intracereroventricular day 6 in the Anti-ANG-(1-12) IgG group, accompanied by higher plasma osmolality on day 13, but there were no differences in urine volume, food intake, or body weight during the 2-wk treatment. In Preimmune IgG-treated animals, there were no significant changes in these variables over the 2-wk period. The antihypertensive effects produced by endogenous neutralization of brain ANG-(1-12) suggest that ANG-(1-12) is functionally active in brain pathways regulating blood pressure.

Isa, Katsunori; Garcia-Espinosa, Maria Antonia; Arnold, Amy C.; Pirro, Nancy T.; Tommasi, Ellen N.; Ganten, Detlev; Chappell, Mark C.; Ferrario, Carlos M.; Diz, Debra I.

2009-01-01

179

Directed expression of transgenes to alveolar type I cells in the mouse.  

Science.gov (United States)

Podoplanin (RTI40, aggrus, T1alpha, hT1alpha-2, E11, PA2.26, RANDAM-2, gp36, gp38, gp40, OTS8) is a type I cell marker in rat lung. We show that a bacterial artificial chromosome vector containing the rat podoplanin gene (RTIbac) delivers a pattern of transgene expression in lung that is more restricted to mouse type I cells than that of the endogenous mouse podoplanin gene. RTIbac-transgenic mice expressed rat podoplanin in type I cells; type II cells, airways, and vascular endothelium were negative. A modified bacterial artificial chromosome containing internal ribosome entry site (IRES)-green fluorescent protein (GFP) sequences in the podoplanin 3'UTR expressed rat podoplanin and transgenic GFP in type I cells. RTIbac transgene expression was absent or reduced in pulmonary pleura, lymphatic endothelium, and putative lymphoid-associated stromal tissue, all of which contained abundant mouse podoplanin. Rat podoplanin mRNA levels in normal rat lung and RTIbac transgenic lung were 25-fold higher than in corresponding kidney and brain samples. On Western blots, transgenic rat and endogenous mouse podoplanin displayed very similar patterns of protein expression in various organs. Highest protein levels were observed in lung with 10- to 20-fold less in brain; there were low levels in thymus and kidney. Both GFP and rat podoplanin transgenes were expressed at extrapulmonary sites of endogenous mouse podoplanin gene expression, including choroid plexus, eye ciliary epithelium, and renal glomerulus. Because their pulmonary expression is more restricted than endogenous mouse podoplanin, RTIbac derivatives should be useful for mouse type I cell-specific transgene delivery. PMID:18367724

Vanderbilt, Jeff N; Allen, Lennell; Gonzalez, Robert F; Tigue, Zachary; Edmondson, Jess; Ansaldi, Daniel; Gillespie, Anne Marie; Dobbs, Leland G

2008-09-01

180

Transgenic mice susceptible to poliovirus.  

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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

 
 
 
 
181

Hepatic fibrosis, glomerulosclerosis, and a lipodystrophy-like syndrome in PEPCK-TGF-beta1 transgenic mice.  

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Transgenic mice overexpressing a constitutively active human TGF-beta1 under control of the rat phosphoenolpyruvate carboxykinase regulatory sequences developed fibrosis of the liver, kidney, and adipose tissue, and exhibited a severe reduction in body fat. Expression of the transgene in hepatocytes resulted in increased collagen deposition, altered lobular organization, increased hepatocyte turnover, and in extreme cases, hemorrhage and thrombosis. Renal expression of the transgene was local...

Clouthier, D. E.; Comerford, S. A.; Hammer, R. E.

1997-01-01

182

Directed Expression of Transgenes to Alveolar Type I Cells in the Mouse  

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Podoplanin (RTI40, aggrus, T1?, hT1?-2, E11, PA2.26, RANDAM-2, gp36, gp38, gp40, OTS8) is a type I cell marker in rat lung. We show that a bacterial artificial chromosome vector containing the rat podoplanin gene (RTIbac) delivers a pattern of transgene expression in lung that is more restricted to mouse type I cells than that of the endogenous mouse podoplanin gene. RTIbac-transgenic mice expressed rat podoplanin in type I cells; type II cells, airways, and vascular endothelium were negati...

Vanderbilt, Jeff N.; Allen, Lennell; Gonzalez, Robert F.; Tigue, Zachary; Edmondson, Jess; Ansaldi, Daniel; Gillespie, Anne Marie; Dobbs, Leland G.

2008-01-01

183

Pharmacogenetic heterogeneity of transgene expression in muscle and tumours  

Directory of Open Access Journals (Sweden)

Full Text Available 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 on the transcription efficiency of the transgene. In the present paper we investigated if transfection of muscle and tumour tissue were also subject to such variations. Methods Variation, in transgene expression, after intramuscular gene delivery was determined in different rodent strains and gene expression in tumours was investigated in different human and rodent cell lines as well as in subcutaneously implanted rodent tumours. The molecular mechanisms involved in transgene expression were dissected using an adenovirus encoding luciferase. The luciferase activity, the viral DNA copies and the luciferase transcripts were assessed in cultured cells as well as in the tissues. Results Large differences of luciferase activity, up to 2 logs, were observed between different rodent strains after intramuscular injection of Ad Luciferase. This inter-strain variation of transgene expression was due to a difference in transcription efficiency. The transgene expression level in tumour cell lines of different tissue origin could be explained largely by the difference of infectibility to the adenovirus. In contrast, the main step responsible for luciferase activity variation, between six human breast cancer cell lines with similar phenotype, was at the transcriptional level. Conclusion Difference in transcriptional efficiency in muscles as observed between different inbred strains and between human breast cancer cell lines may be expected to occur between individual patients. This might have important consequences for clinical gene therapy. The variation between tumour types and tissues within a species are mainly at the levels of infectivity.

Attema Joline

2003-08-01

184

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

185

Specific transgene expression in mouse pancreatic ?-cells under the control of the porcine insulin promoter  

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Abstract The availability of regulatory sequences directing tissue-specific expression of transgenes in genetically modified mice and large animals is a prerequisite for the development of adequate models for human diseases. The rat insulin 2 gene (Ins2) promoter, widely used to achieve transgene expression in pancreatic ?-cells of mice, also directs expression to extrapancreatic tissues and performs poorly in isolated pancreatic islets of human, mouse, and pig. To evaluate whethe...

2009-01-01

186

Development of transgenic animals for optogenetic manipulation of mammalian nervous system function: progress and prospects for behavioral neuroscience.  

Science.gov (United States)

Here we review the rapidly growing toolbox of transgenic mice and rats that exhibit functional expression of engineered opsins for neuronal activation and silencing with light. Collectively, these transgenic animals are enabling neuroscientists to access and manipulate the many diverse cell types in the mammalian nervous system in order to probe synaptic and circuitry connectivity, function, and dysfunction. The availability of transgenic lines affords important advantages such as stable and heritable transgene expression patterns across experimental cohorts. As such, the use of transgenic lines precludes the need for other costly and labor-intensive procedures to achieve functional transgene expression in each individual experimental animal. This represents an important consideration when large cohorts of experimental animals are desirable as in many common behavioral assays. We describe the diverse strategies that have been implemented for developing transgenic mouse and rat lines and highlight recent advances that have led to dramatic improvements in achieving functional transgene expression of engineered opsins. Furthermore, we discuss considerations and caveats associated with implementing recently developed transgenic lines for optogenetics-based experimentation. Lastly, we propose strategies that can be implemented to develop and refine the next generation of genetically modified animals for behaviorally-focused optogenetics-based applications. PMID:23473879

Ting, Jonathan T; Feng, Guoping

2013-10-15

187

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. 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.)

1993-11-01

188

Transgenic mice in developmental toxicology.  

Science.gov (United States)

Advances in molecular biology and embryology are being utilized for the generation of transgenic mice, animals that contain specific additions, deletions, or modifications of genes or sequences in their DNA. Mouse embryonic stem cells and homologous recom...

R. P. Woychik

1992-01-01

189

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

190

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)

2010-02-01

191

Transgene Expression Is Associated with Copy Number and Cytomegalovirus Promoter Methylation in Transgenic Pigs  

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Transgenic animals have been used for years to study gene function, produce important proteins, and generate models for the study of human diseases. However, inheritance and expression instability of the transgene in transgenic animals is a major limitation. Copy number and promoter methylation are known to regulate gene expression, but no report has systematically examined their effect on transgene expression. In the study, we generated two transgenic pigs by somatic cell nuclear transfer (S...

Kong, Qingran; Wu, Meiling; Huan, Yanjun; Zhang, Li; Liu, Haiyan; Bou, Gerelchimeg; Luo, Yibo; Mu, Yanshuang; Liu, Zhonghua

2009-01-01

192

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

193

The last intron of the human thrombopoietin gene enhances expression in milk of transgenic mice.  

Science.gov (United States)

Introns can enhance gene expression levels. This effect is known as intron-mediated enhancement, which is different from that of enhancers or promoters. In our previous study, under the control of the cytomegalovirus or goat ?-casein promoter, the vector containing intron V-TPOcDNA expressed the highest thrombopoietin (TPO) level, whereas the vector containing TPOgDNA expressed the lowest level. In order to verify whether intron V also improves TPO expression in the milk of transgenic mice, rat whey acidic protein promoter was used as regulatory element to construct mammary gland expression vectors including pTPOWA (containing TPOcDNA), pTPOWB (containing intron V-TPOcDNA), and pTPOWC (containing TPOgDNA). These vectors were transfected into HC-11 cells and the supernatants were analyzed at 48 h. The highest TPO level was found in pTPOWB (795 pg/mL) and the lowest level in pTPOWC (193 pg/mL). Then, corresponding vectors were microinjected into fertilized mice zygotes. Transgenic mice were identified by polymerase chain reaction and Southern blot. Enzyme-linked immunosorbent assay was performed to measure TPO levels in the milk of lactating transgenic mice. The highest and lowest TPO levels were found in transgenic mice carrying intron V-TPOcDNA (2,307 pg/mL) and in transgenic mice carrying TPOgDNA (242 pg/mL), respectively. Thus, intron V remarkably improved TPO expression in transgenic mice. PMID:24287579

Li, Yan; Zhou, Mingqian; Zhou, Hongwei; Ning, Yunshan

2014-03-01

194

Transposon-mediated transgenesis, transgenic rescue, and tissue-specific gene expression in rodents and rabbits.  

Science.gov (United States)

Germline transgenesis is an important procedure for functional investigation of biological pathways, as well as for animal biotechnology. We have established a simple, nonviral protocol in three important biomedical model organisms frequently used in physiological studies. The protocol is based on the hyperactive Sleeping Beauty transposon system, SB100X, which reproducibly promoted generation of transgenic founders at frequencies of 50-64, 14-72, and 15% in mice, rats, and rabbits, respectively. The SB100X-mediated transgene integrations are less prone to genetic mosaicism and gene silencing as compared to either the classical pronuclear injection or to lentivirus-mediated transgenesis. The method was successfully applied to a variety of transgenes and animal models, and can be used to generate founders with single-copy integrations. The transposon vector also allows the generation of transgenic lines with tissue-specific expression patterns specified by promoter elements of choice, exemplified by a rat reporter strain useful for tracking serotonergic neurons. As a proof of principle, we rescued an inborn genetic defect in the fawn-hooded hypertensive rat by SB100X transgenesis. A side-by-side comparison of the SB100X- and piggyBac-based protocols revealed that the two systems are complementary, offering new opportunities in genome manipulation. PMID:23195032

Katter, Katharina; Geurts, Aron M; Hoffmann, Orsolya; Mátés, Lajos; Landa, Vladimir; Hiripi, László; Moreno, Carol; Lazar, Jozef; Bashir, Sanum; Zidek, Vaclav; Popova, Elena; Jerchow, Boris; Becker, Katja; Devaraj, Anantharam; Walter, Ingrid; Grzybowksi, Michael; Corbett, Molly; Filho, Artur Rangel; Hodges, Matthew R; Bader, Michael; Ivics, Zoltán; Jacob, Howard J; Pravenec, Michal; Bosze, Zsuzsanna; Rülicke, Thomas; Izsvák, Zsuzsanna

2013-03-01

195

An inbred colony of oncogene transgenic mice: diversity of tumours and potential as a therapeutic model.  

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Transgenic mice carrying the activated rat c-neu oncogene under transcriptional control of the MMTV promoter were backcrossed to BALB/c mice, with the aim of developing a model for cancer therapy. A total of 86 of 268 transgene-positive mice in the first five generations developed 93 histologically diverse tumours (median age of onset 18 months). The cumulative incidence of breast tumours at 24 months was 18%, and overall tumour incidence 31%. As well as expected c-neu expressing breast cance...

Thomas, H.; Hanby, A. M.; Smith, R. A.; Hagger, P.; Patel, K.; Raikundalia, B.; Camplejohn, R. S.; Balkwill, F. R.

1996-01-01

196

Transgenic Zebrafish Using Transposable Elements  

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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

197

Transgenic models of Huntington's disease.  

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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

198

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.

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

199

Expression Systems and Species Used for Transgenic Animal Bioreactors  

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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 ...

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

2013-01-01

200

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

 
 
 
 
201

Identification of the Transgenic Integration Site in Immunodeficient tg?26 Human CD3? Transgenic Mice  

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A strain of human CD3? transgenic mice, tg?26, exhibits severe immunodeficiency associated with early arrest of T cell development. Complete loss of T cells is observed in homozygous tg?26 mice, but not in heterozygotes, suggesting that genomic disruption due to transgenic integration may contribute to the arrest of T cell development. Here we report the identification of the transgenic integration site in tg?26 mice. We found that multiple copies of the human CD3? transgene are inserted...

Ohigashi, Izumi; Yamasaki, Yuki; Hirashima, Tsukasa; Takahama, Yousuke

2010-01-01

202

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

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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

203

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

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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

204

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

2003-06-20

205

Transgenic Spartina alterniflora for phytoremediation.  

Science.gov (United States)

Perennial monoculture forming grasses are very important natural remediators of pollutants. Their genetic improvement is an important task because introduction of key transgenes can dramatically improve their remediation potential. Transfer of key genes for mercury phytoremediation into the salt marsh cordgrass (Spartina alterniflora) is reported here. S. alterniflora plays an important role in the salt marsh by cycling of elements, both nutrients and pollutants, protects the coastline from erosion, is a keystone species in the salt marsh supporting a large food web, which in turn supports a significant segment of economy, including tourism, has an impact on cloud formation and consequently on global weather, and is thus an ecologically important species relevant for our life-support systems. Embryogenic callus of S. alterniflora was co-inoculated with a pair of Agrobacterium strains LBA4404 carrying the organomercurial lyase (merB) and mercuric reductase (merA) genes, respectively, in order to co-introduce both the merA and the merB genes. Seven stable geneticin resistant lines were recovered. The presence of merA and merB genes was verified by PCR and Southern blotting. All but one transgenic lines contained both the merA and the merB sequences proving that co-introduction into Spartina of two genes from separate Agrobacterium strains is feasible and frequent, although the overall frequency of transformation is low. Northern blotting showed differences in relative expression of the two transgenes among individual transformants. The steady-state RNA levels appeared to correlate with the phenotype. Line #7 showed the highest resistance to HgCl(2) (up to 500 microM), whereas line #3 was the most resistant to phenylmercuric acetate (PMA). Wild-type (WT) callus is sensitive to PMA at 50 microM and to HgCl(2) at 225 microM. PMID:16528587

Czakó, Mihály; Feng, Xianzhong; He, Yuke; Liang, Dali; Márton, László

2006-01-01

206

Improving expression of reporter transgene in stem cell by construction of different lentiviral vectors  

International Nuclear Information System (INIS)

For stem cell trafficking applications, it is imperative to express transgenes at desired and stable levels. In recent years, lentivirus-mediated gene transfer was shown to be an efficient method to stably introduce genetic modifications in target cells, even if these are in proliferative or nonproliferative states. Moreover, transgene expression levels can be controlled by using different promoters. The present study was designed to compare the potency of various promoters regulating expression of imaging reporter genes in embryonic H9c2 cardiomyoblasts derived from rat heart. Lentiviral vector was produced by the transient transfection of plasmids carrying required genes and those encoding for virus coating proteins into 293T cells. Harvested viral constructs were incubated with Hela and H9c2 cells, respectively. Transgene expressions were detected by several imaging modalities and evaluated by enzymatic assays. Results - We observed that the level of stable transgene expression in lentivirus-transduced myoblasts could be modulated over several orders of magnitude, with the Ubiquitin (Ub) promoter exhibiting the highest activity, intermediate expression was observed with the CAG promoter, whereas expression observed with the CMV promoter was very weak. We observed that the level of stable transgene expression in lentivirus-transduced myoblasts could be modulated over several orders of magnitude, with the Ubiquitin (Ub) promoter exhibiting the highest activity, intermediate expression was observed with the CAG promoter, whereas expression observed with the CMV promoter was very weak. Here we show that lentivirus-mediated gene transfer allows efficient and stable transgene expression in embryonic cardiomyoblasts in vitro and that transgene expression levels can be varied by using different well-characterized gene promoters. In vivo trials about gene expression will probably further determine the potential of long-term trafficking stem cells using lentivirus

2007-10-26

207

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

208

High expression of transgene protein in Spirodela.  

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The monocot family Lemnaceae (duckweed) is composed of small, edible, aquatic plants. Spirodela oligorrhiza SP is a duckweed with a biomass doubling time of about 2 days under controlled, axenic conditions. Stably transformed Spirodela plants were obtained following co-cultivation of regenerative calli with Agrobacterium tumefaciens. GFP activity was successfully monitored in different subcellular compartments of the plant and correlated with different targeting sequences. Transgenic lines were followed for a period of at least 18 months and more than 180 vegetative doublings (generations). The lines are stable in morphology, growth rate, transgene expression, and activity as measured by DNA-DNA and immunoblot hybridizations, fluorescence activity measurements, and antibiotic resistance. The level of transgene expression is a function of leader sequences rather than transgene copy number. A stable, transgenic, GFP expression level >25% of total soluble protein is demonstrated for the S. oligorrhiza system, making it among the higher expressing systems for nuclear transformation in a higher plant. PMID:17492286

Vunsh, Ron; Li, Jihong; Hanania, Uri; Edelman, Marvin; Flaishman, Moshe; Perl, Avihai; Wisniewski, Jean-Pierre; Freyssinet, Georges

2007-09-01

209

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 (TF<1) at all levels of metal treatment. Among the 4 transgenic 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

210

Generation of NSE-MerCreMer Transgenic Mice with Tamoxifen Inducible Cre Activity in Neurons  

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To establish a genetic tool for conditional deletion or expression of gene in neurons in a temporally controlled manner, we generated a transgenic mouse (NSE-MerCreMer), which expressed a tamoxifen inducible type of Cre recombinase specifically in neurons. The tamoxifen inducible Cre recombinase (MerCreMer) is a fusion protein containing Cre recombinase with two modified estrogen receptor ligand binding domains at both ends, and is driven by the neural-specific rat neural specific enolase (NS...

Kam, Mandy Ka Man; Lee, King Yiu; Tam, Paul Kwong Hang; Lui, Vincent Chi Hang

2012-01-01

211

Malignant Progression and Blockade of Angiogenesis in a Murine Transgenic Model of Neuroblastoma  

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Targeted expression of MYCN to the neural crest [under control of the rat tyrosine hydroxylase (TH) promoter] causes neuroblastoma in transgenic mice (TH-MYCN) and is a well-established model for this disease. Because high levels of MYCN are associated with enhanced tumor angiogenesis and poor clinical outcome in neuroblastoma, we serially characterized malignant progression, angiogenesis, and sensitivity to angiogenic blockade in tumors from these animals. Tumor cells were proliferative, sec...

2007-01-01

212

Transgenic overexpression of PKC? in the mouse prostate induces preneoplastic lesions  

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It is well established that protein kinase C (PKC) isozymes play distinctive roles in mitogenic and survival signaling as well as in cancer progression. PKC?, the product of the PRKCE gene, is upregulated in various types of cancers including prostate, lung and breast cancer. To address a potential role for PKCs in prostate cancer progression we generated three mouse transgenic lines expressing PKC?, PKC? or PKC? in the prostate epithelium under the control of the rat probasin (PB) promot...

Benavides, Fernando; Blando, Jorge; Perez, Carlos J.; Garg, Rachana; Conti, Claudio J.; Digiovanni, John; Kazanietz, Marcelo G.

2011-01-01

213

G2R Cre Reporter Transgenic Zebrafish  

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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

214

Cardiac phenotype induced by a dysfunctional ?1C transgene: A general problem for the transgenic approach  

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Based on stable integration of recombinant DNA into a host genome, transgenic technology has become an important genetic engineering methodology. An organism whose genetic characteristics have been altered by the insertion of foreign DNA is supposed to exhibit a new phenotype associated with the function of the transgene. However, successful insertion may not be sufficient to achieve specific modification of function. In this study we describe a strain of transgenic mouse, G7-882, generated b...

Asemu, Girma; Fishbein, Kenneth; Lao, Qi Zong; Ravindran, Arippa; Herbert, Ron; Canuto, Holly C.; Spencer, Richard G.; Soldatov, Nikolai M.

2011-01-01

215

Quantitative real-time polymerase chain reaction (qRT-PCR) restriction fragment length polymorphism (RFLP) method for monitoring highly conserved transgene expression during gene therapy.  

Science.gov (United States)

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 quantitative real-time polymerase chain reaction (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 vitro, murine AML12 hepatocytes were transfected with rat HO-1. After 40 h, the total HO-1 mRNA was enriched 2-fold relative to control cells, and rat HO-1 comprised 84% of HO-1 cDNA. In vivo, the rat HO-1 transgene was cloned into a Sleeping Beauty transposase (SB-Tn) construct and was injected hydrodynamically into a mouse model of sickle cell disease (SCD). After 21 days, there was a 32% enrichment of HO-1 mRNA relative to control mice and the rat transgene comprised 88% of HO-1 cDNA. After 21 days, HO-1 protein expression in liver was increased 2.5-fold. In summary, qRT-PCR RFLP is a useful and reliable method to differentiate the transgene from host gene transcription, especially when the host and transgene protein are identical or highly homologous. This method has translational applications to the design, delivery, and monitoring of gene-therapy vectors. PMID:19059164

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

2008-12-01

216

Wheat storage proteins in transgenic rice endosperm.  

Science.gov (United States)

Transgenic rice seed expressing wheat HMW glutenin subunit was characterized to study the effects of the wheat prolamin on the protein expression pattern and protein size distribution in the endosperm and the functional and rheological properties of the rice flour and dough. Significant differences were found in the protein expression pattern between the transgenic and wild type samples. Comparing the protein expression profiles of transgenic and nontransgenic plants, combined with proteomic-based studies, indicated increased protein disulfide isomerase (PDI) levels in the transgenic rice lines. The accurate molecular size of HMW-GS in rice endosperm was identified by MALDI-TOF-MS analysis. The expressed wheat HMW (subunit 1Dx5) GS showed a positive effect on the functional properties of rice dough by significantly increasing the size distribution of the polymeric protein fraction and modifying the dough mixing parameters. PMID:23802557

Oszvald, Mária; Balázs, Gábor; Pólya, Sára; Tömösközi, Sándor; Appels, Rudi; Békés, Ferenc; Tamás, László

2013-08-01

217

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.

218

Pollen selection: a transgenic reconstruction approach.  

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A transgenic reconstruction experiment has been performed to determine the feasibility of male gametophytic selection to enhance transmission of genes to the next sporophytic generation. For tobacco pollen from a transgenic plant containing a single hygromycin-resistance (hygromycin phosphotransferase, hpt-) gene under control of the dc3 promoter, which is active in both sporophytic and gametophytic tissues, 3 days of in vitro maturation in hygromycin-containing medium was sufficient to resul...

Touraev, A.; Fink, C. S.; Sto?ger, E.; Heberle-bors, E.

1995-01-01

219

Involution of the lactating mammary gland is inhibited by the IGF system in a transgenic mouse model.  

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Development of the mammary gland during puberty, pregnancy, and lactation is controlled by steroid and peptide hormones and growth factors. To determine the role of the insulin-like growth factors (IGFs) in this process we developed a transgenic model using the whey acidic protein (WAP) gene to direct expression of rat IGF-I and human IGF binding protein-3 (IGFBP-3) to mammary tissue during late pregnancy and throughout lactation. High levels of expression of transgenic IGF-I and IGFBP-3 were...

Neuenschwander, S.; Schwartz, A.; Wood, T. L.; Roberts, C. T.; Hennighausen, L.; Leroith, D.

1996-01-01

220

Selenoprotein-Transgenic Chlamydomonas reinhardtii  

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Full Text Available Selenium (Se deficiency is associated with the occurrence of many diseases. However, excessive Se supplementation, especially with inorganic Se, can result in toxicity. Selenoproteins are the major forms of Se in vivo to exert its biological function. Expression of those selenoproteins, especially with the application of a newly developed system, is thus very important for studying the mechanism of Se in nutrition. The use of Chlamydomonas reinhardtii (C. reinhardtii as a biological vector to express an heterogeneous protein is still at the initial stages of development. In order to investigate the possibility of using this system to express selenoproteins, human 15-KDa selenoprotein (Sep15, a small but widely distributed selenoprotein in mammals, was chosen for the expression platform test. Apart from the wild-type human Sep15 gene fragment, two Sep15 recombinants were constructed containing Sep15 open reading frame (ORF and the selenocysteine insertion sequence (SECIS element from either human Sep15 or C. reinhardtii selenoprotein W1, a highly expressed selenoprotein in this alga. Those Sep15-containing plasmids were transformed into C. reinhardtii CC-849 cells. Results showed that Sep15 fragments were successfully inserted into the nuclear genome and expressed Sep15 protein in the cells. The transgenic and wild-type algae demonstrated similar growth curves in low Se culture medium. To our knowledge, this is the first report on expressing human selenoprotein in green alga.

Jiazuan Ni

2013-02-01

 
 
 
 
221

Transgenic technologies to induce sterility  

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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

222

A Primer for Using Transgenic Insecticidal Cotton in Developing Countries  

Science.gov (United States)

Many developing countries face the decision of whether to approve the testing and commercial use of insecticidal transgenic cotton and the task of developing adequate regulations for its use. In this review, we outline concepts and provide information to assist farmers, regulators and scientists in making decisions concerning this technology. We address seven critical topics: 1) molecular and breeding techniques used for the development of transgenic cotton cultivars, 2) properties of transgenic cotton cultivars and their efficacy against major insect pests, 3) agronomic performance of transgenic cotton in developing countries, 4) factors affecting transgene expression, 5) impact of gene flow between transgenic and non-transgenic cotton, 6) non-target effects of transgenic cotton, and 7) management of pest resistance to transgenic cotton.

Showalter, Ann M.; Heuberger, Shannon; Tabashnik, Bruce E.; Carriere, Yves

2009-01-01

223

Growth Factor Transgenes Interactively Regulate Articular Chondrocytes  

Science.gov (United States)

Adult articular chondrocytes lack an effective repair response to correct damage from injury or osteoarthritis. Polypeptide growth factors that stimulate articular chondrocyte proliferation and cartilage matrix synthesis may augment this response. Gene transfer is a promising approach to delivering such factors. No single growth factor gene is likely to optimize these cell functions, but multiple growth factor gene transfer remains unexplored. We tested the hypothesis that multiple growth factor gene transfer selectively modulates articular chondrocyte proliferation and matrix synthesis. We tested the hypothesis by delivering combinations of the transgenes encoding insulin-like growth factor I (IGF-I), fibroblast growth factor-2 (FGF-2), transforming growth factor beta1 (TGF-?1), bone morphogenetic protein-2 (BMP-2) and bone morphogenetic protien-7 (BMP-7) to articular chondrocytes and measured changes in the production of DNA, glycosaminoglycan and collagen. The transgenes differentially regulated all these chondrocyte functions. In concert, the transgenes interacted to generate widely divergent responses from the cells. These interactions ranged from inhibitory to synergistic. The transgene pair encoding IGF-I and FGF-2 maximized cell proliferation. The three-transgene group encoding IGF-I, BMP-2 and BMP-7 maximized matrix production and also optimized the balance between cell proliferation and matrix production. These data demonstrate a potentially tunable approach to articular chondrocyte regulation and suggest that certain growth factor gene combinations have potential value for cell-based articular cartilage repair.

Shi, Shuiliang; Mercer, Scott; Eckert, George J.; Trippel, Stephen B.

2014-01-01

224

Growth factor transgenes interactively regulate articular chondrocytes.  

Science.gov (United States)

Adult articular chondrocytes lack an effective repair response to correct damage from injury or osteoarthritis. Polypeptide growth factors that stimulate articular chondrocyte proliferation and cartilage matrix synthesis may augment this response. Gene transfer is a promising approach to delivering such factors. Multiple growth factor genes regulate these cell functions, but multiple growth factor gene transfer remains unexplored. We tested the hypothesis that multiple growth factor gene transfer selectively modulates articular chondrocyte proliferation and matrix synthesis. We tested the hypothesis by delivering combinations of the transgenes encoding insulin-like growth factor I (IGF-I), fibroblast growth factor-2 (FGF-2), transforming growth factor beta1 (TGF-?1), bone morphogenetic protein-2 (BMP-2), and bone morphogenetic protien-7 (BMP-7) to articular chondrocytes and measured changes in the production of DNA, glycosaminoglycan, and collagen. The transgenes differentially regulated all these chondrocyte activities. In concert, the transgenes interacted to generate widely divergent responses from the cells. These interactions ranged from inhibitory to synergistic. The transgene pair encoding IGF-I and FGF-2 maximized cell proliferation. The three-transgene group encoding IGF-I, BMP-2, and BMP-7 maximized matrix production and also optimized the balance between cell proliferation and matrix production. These data demonstrate an approach to articular chondrocyte regulation that may be tailored to stimulate specific cell functions, and suggest that certain growth factor gene combinations have potential value for cell-based articular cartilage repair. PMID:23097312

Shi, Shuiliang; Mercer, Scott; Eckert, George J; Trippel, Stephen B

2013-04-01

225

Induction of melanoma in TPras transgenic mice.  

Science.gov (United States)

In order to study the oncogenesis of melanocytes, transgenic mouse lines were established that express a mutated human Ha-ras (TPras) gene in pigment producing cells. The ras transgenic mice exhibit an altered phenotype, including melanocytic hyperplasia and a muted agouti coat, indicative of hyperproliferative melanocytes. These mice and their wild-type littermates have been subjected to a variety of carcinogenesis protocols, including 7, 12-dimethylbenz-[a]anthracene (DMBA), 12-O-tetradecanoylphorbol-13-acetate (TPA) and UV radiation exposure. Topical DMBA treatment of TPras mice resulted in a high incidence of melanomas. Metastatic lesions were observed in skin, lungs and lymph nodes. TPA treatment of TPras mice induced a small number of papillomas but no nevi or melanomas. UV light exposures induced papillomas in negative littermate and melanomas in some albino TPras mice. These results show that melanocytes expressing an activated Ha-ras in the TPras transgenic mice are susceptible to induction of melanoma by DMBA. PMID:10469620

Broome Powell, M; Gause, P R; Hyman, P; Gregus, J; Lluria-Prevatt, M; Nagle, R; Bowden, G T

1999-09-01

226

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

227

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

228

Transgenic Rescue of Enamel Phenotype in Ambn Null Mice  

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Ameloblastin null mice fail to make an enamel layer, but the defects could be due to an absence of functional ameloblastin or to the secretion of a potentially toxic mutant ameloblastin. We hypothesized that the enamel phenotype could be rescued by the transgenic expression of normal ameloblastin in Ambn mutant mice. We established and analyzed 5 transgenic lines that expressed ameloblastin from the amelogenin (AmelX) promoter and identified transgenic lines that express virtually no transgen...

2010-01-01

229

Detection of potential transgenic plant DNA recipients among soil bacteria  

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The likelihood of gene transfer from transgenic plants to bacteria is dependent on gene number and the presence of homologous sequences. The large number of transgene copies in transplastomic (transgenes contained in the chloroplast genome) plant cells as well as the prokaryotic origin of the transgene, may thus significantly increase the likelihood of gene transfer to bacteria that colonize plant tissues. In order to assess the probability of such transfer, the length of homologous DNA seque...

Monier, Jean-michel; Bernillon, Dominique; Kay, Elizabeth; Faugier, Aure?lie; Rybalka, Oleksandra; Dessaux, Yves; Simonet, Pascal; Vogel, Timothy

2007-01-01

230

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

231

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; 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.

2009-09-03

232

Combined Allogeneic Tumor Cell Vaccination and Systemic Interleukin 12 Prevents Mammary Carcinogenesis in HER-2/neu Transgenic Mice  

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Transgenic Balb/c mice expressing the transforming rat HER-2/neu oncogene develop early and multifocal mammary carcinomas. Within the first 5 months of life the tissue-specific expression of HER-2/neu causes a progression in all their 10 mammary glands from atypical hyperplasia to invasive carcinoma. It was previously observed that chronic administration of interleukin (IL)-12 increased tumor latency, but every mouse eventually succumbed to multiple carcinomas. A significant improvement in tu...

Nanni, Patrizia; Nicoletti, Giordano; Giovanni, Carla; Landuzzi, Lorena; Di Carlo, Emma; Cavallo, Federica; Pupa, Serenella M.; Rossi, Ilaria; Colombo, Mario P.; Ricci, Cinzia; Astolfi, Annalisa; Musiani, Piero; Forni, Guido; Lollini, Pier-luigi

2001-01-01

233

Combined allogeneic tumor cell vaccination and systemic interleukin 12 prevents mammary carcinogenesis in HER-2/neu transgenic mice.  

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Transgenic Balb/c mice expressing the transforming rat HER-2/neu oncogene develop early and multifocal mammary carcinomas. Within the first 5 months of life the tissue-specific expression of HER-2/neu causes a progression in all their 10 mammary glands from atypical hyperplasia to invasive carcinoma. It was previously observed that chronic administration of interleukin (IL)-12 increased tumor latency, but every mouse eventually succumbed to multiple carcinomas. A significant improvement in tu...

Forni, Guido; Cavallo, Federica

2001-01-01

234

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 ...

235

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 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

1994-05-07

236

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

237

Generation of transgenic Xenopus using restriction enzyme-mediated integration.  

Science.gov (United States)

Transgenesis, the process of incorporating an exogenous gene (transgene) into an organism's genome, is a widely used tool to develop models of human diseases and to study the function and/or regulation of genes. Generating transgenic Xenopus is rapid and involves simple in vitro manipulations, taking advantage of the large size of the amphibian egg and external embryonic development. Restriction enzyme-mediated integration (REMI) has a number of advantages for transgenesis compared to other methods used to produce transgenic Xenopus, including relative efficiency, higher transgene expression levels, fewer genetic chimera in founder transgenic animals, and near-complete germ-line transgene transmission. This chapter explains the REMI method for generating transgenic Xenopus laevis tadpoles, including improvements developed to enable studies in the mature retina. PMID:22688696

Haeri, Mohammad; Knox, Barry E

2012-01-01

238

Insertional mutagenesis by a hybrid piggyBac and sleeping beauty transposon in the rat.  

Science.gov (United States)

A hybrid piggyBac/Sleeping Beauty transposon-based insertional mutagenesis system that can be mobilized by simple breeding was established in the rat. These transposons were engineered to include gene trap sequences and a tyrosinase (Tyr) pigmentation reporter to rescue the albinism of the genetic background used in the mutagenesis strategy. Single-copy transposon insertions were transposed into the rat genome by co-injection of plasmids carrying the transposon and RNA encoding piggyBac transposase into zygotes. The levels of transgenic Tyr expression were influenced by chromosomal context, leading to transgenic rats with different pigmentation that enabled visual genotyping. Transgenic rats designed to ubiquitously express either piggyBac or Sleeping Beauty transposase were generated by standard zygote injection also on an albino background. Bigenic rats carrying single-copy transposons at known loci and transposase transgenes exhibited coat color mosaicism, indicating somatic transposition. PiggyBac or Sleeping Beauty transposase bigenic rats bred with wild-type albino rats yielded offspring with pigmentation distinct from the initial transposon insertions as a consequence of germline transposition to new loci. The germline transposition frequency for Sleeping Beauty and piggyBac was ?10% or about one new insertion per litter. Approximately 50% of the insertions occurred in introns. Chimeric transcripts containing endogenous and gene trap sequences were identified in Gabrb1 mutant rats. This mutagenesis system based on simple crosses and visual genotyping can be used to generate a collection of single-gene mutations in the rat. PMID:23023007

Furushima, Kenryo; Jang, Chuan-Wei; Chen, Diane W; Xiao, Ningna; Overbeek, Paul A; Behringer, Richard R

2012-12-01

239

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...

2004-01-01

240

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

 
 
 
 
241

Protective effects of transgenic human endothelial protein C receptor expression in murine models of transplantation.  

Science.gov (United States)

Thrombosis and inflammation are major obstacles to successful pig-to-human solid organ xenotransplantation. A potential solution is genetic modification of the donor pig to overexpress molecules such as the endothelial protein C receptor (EPCR), which has anticoagulant, anti-inflammatory and cytoprotective signaling properties. Transgenic mice expressing human EPCR (hEPCR) were generated and characterized to test this approach. hEPCR was expressed widely and its compatibility with the mouse protein C pathway was evident from the anticoagulant phenotype of the transgenic mice, which exhibited a prolonged tail bleeding time and resistance to collagen-induced thrombosis. hEPCR mice were protected in a model of warm renal ischemia reperfusion injury compared to wild type (WT) littermates (mean serum creatinine 39.0 ± 2.3 ?mol/L vs. 78.5 ± 10.0 ?mol/L, p < 0.05; mean injury score 31 ± 7% vs. 56 ± 5%, p < 0.05). Heterotopic cardiac xenografts from hEPCR mice showed a small but significant prolongation of survival in C6-deficient PVG rat recipients compared to WT grafts (median graft survival 6 vs. 5 days, p < 0.05), with less hemorrhage and edema in rejected transgenic grafts. These data indicate that it is possible to overexpress EPCR at a sufficient level to provide protection against transplant-related thrombotic and inflammatory injury, without detrimental effects in the donor animal. PMID:22681753

Lee, K F E; Lu, B; Roussel, J C; Murray-Segal, L J; Salvaris, E J; Hodgkinson, S J; Hall, B M; d'Apice, A J F; Cowan, P J; Gock, H

2012-09-01

242

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)

2006-03-01

243

Transgene mobilization and regulatory uncertainty for non-GE fruit products of transgenic rootstocks.  

Science.gov (United States)

Genetically engineered (GE) rootstocks may offer some advantages for biotechnology applications especially in woody perennial crops such as grape or walnut. Transgrafting combines horticultural grafting practices with modern GE methods for crop improvement. Here, a non-GE conventional scion (upper stem portion) is grafted onto a transgenic GE rootstock. Thus, the scion does not contain the genetic modification present in the rootstock genome. We examined transgene presence in walnut and tomato GE rootstocks and non-GE fruit-bearing scions. Mobilization of transgene DNA, protein, and mRNA across the graft was not detected. Though transgenic siRNA mobilization was not observed in grafted tomatoes or walnut scions, transgenic siRNA signal was detected in walnut kernels. Prospective benefits from transgrafted plants include minimized risk of GE pollen flow (Lev-Yadun and Sederoff, 2001), possible use of more than one scion per approved GE rootstock which could help curb the estimated US$136 million (CropLife International, 2011) cost to bring a GE crop to international markets, as well as potential for improved consumer and market acceptance since the consumable product is not itself GE. Thus, transgrafting provides an alternative option for agricultural industries wishing to expand their biotechnology portfolio. PMID:22749907

Haroldsen, Victor M; Chi-Ham, Cecilia L; Bennett, Alan B

2012-10-31

244

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

245

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) has provided effective ...

Chougule, Nanasaheb P.; Bonning, Bryony C.

2012-01-01

246

Transgenic pig carrying green fluorescent proteasomes  

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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

247

Transgenic rabbits expressing human lipoprotein lipase  

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To study the functions of lipoprotein lipase (LPL) in lipid and lipoprotein metabolism and the relationship between LPL and atherosclerosis, we generated transgenic rabbits expressing the human LPL gene. A total of 4045 Japanese whiterabbit embryos were microinjected with a 3.8-kb SalI/HindIII fragment containing the chicken ?-actin promoter, human LPL cDNA and rabbit ?-globin with poly (A) signals, and then transplanted into 116 recipient rabbits. Of the 166 pups born, six pups were transg...

Araki, Masahiro; Fan, Jianglin; Challah, Mireille; Bensadoun, Andre?; Yamada, Nobuhiro; Honda, Kazuo; Watanabe, Teruo

2000-01-01

248

Toxins for Transgenic Resistance to Hemipteran Pests  

Digital Repository Infrastructure Vision for European Research (DRIVER)

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...

Chougule, Nanasaheb P.; Bonning, Bryony C.

2012-01-01

249

An ovine transgenic Huntington's disease model.  

Science.gov (United States)

Huntington's disease (HD) is an inherited autosomal dominant neurodegenerative disorder caused by an expansion of a CAG trinucleotide repeat in the huntingtin (HTT) gene [Huntington's Disease Collaborative Research Group (1993) A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington's disease chromosomes. The Huntington's Disease Collaborative Research Group. Cell, 72, 971-983]. Despite identification of the gene in 1993, the underlying life-long disease process and effective treatments to prevent or delay it remain elusive. In an effort to fast-track treatment strategies for HD into clinical trials, we have developed a new large-animal HD transgenic ovine model. Sheep, Ovis aries L., were selected because the developmental pattern of the ovine basal ganglia and cortex (the regions primarily affected in HD) is similar to the analogous regions of the human brain. Microinjection of a full-length human HTT cDNA containing 73 polyglutamine repeats under the control of the human promotor resulted in six transgenic founders varying in copy number of the transgene. Analysis of offspring (at 1 and 7 months of age) from one of the founders showed robust expression of the full-length human HTT protein in both CNS and non-CNS tissue. Further, preliminary immunohistochemical analysis demonstrated the organization of the caudate nucleus and putamen and revealed decreased expression of medium size spiny neuron marker DARPP-32 at 7 months of age. It is anticipated that this novel transgenic animal will represent a practical model for drug/clinical trials and surgical interventions especially aimed at delaying or preventing HD initiation. New sequence accession number for ovine HTT mRNA: FJ457100. PMID:20154343

Jacobsen, Jessie C; Bawden, C Simon; Rudiger, Skye R; McLaughlan, Clive J; Reid, Suzanne J; Waldvogel, Henry J; MacDonald, Marcy E; Gusella, James F; Walker, Simon K; Kelly, Jennifer M; Webb, Graham C; Faull, Richard L M; Rees, Mark I; Snell, Russell G

2010-05-15

250

Impaired Fertility in Transgenic Mice Overexpressing Betacellulin.  

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Impaired fertility in transgenic mice overexpressing Betacellulin Peptide growth factors regulate many cellular functions by autocrine, paracrine, juxtacrine or endocrine mechanisms. The epidermal growth factor (EGF)-like peptides are emerging as major players in regulating different aspects of animal and human physiology and pathology. The EGF family elicits essential actions in reproduction. For instance, different Egfr ligands have been shown to be involved in oocyte maturation and ovu...

Gratao, Ana Angelica

2007-01-01

251

Transgenic cereals: current status and future prospects  

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This review summarises the history of transgenic (GM) cereals, principally maize, and then focuses on the scientific literature published in the last two years. It describes the production of GM cereals with modified traits, divided into input traits and output traits. The first category includes herbicide tolerance and insect resistance, and resistance to abiotic and biotic stresses; the second includes altered grains for starch, protein or nutrient quality, the use of cereals for the produc...

2013-01-01

252

Can Transgenic Maize Affect Soil Microbial Communities?  

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The aim of the experiment was to determine if temporal variations of belowground activity reflect the influence of the Cry1Ab protein from transgenic maize on soil bacteria and, hence, on a regulatory change of the microbial community (ability to metabolize sources belonging to different chemical guilds) and/or a change in numerical abundance of their cells. Litter placement is known for its strong influence on the soil decomposer communities. The effects of the addition of crop residues on r...

Mulder, Christian; Wouterse, Marja; Raubuch, Markus; Roelofs, Willem; Rutgers, Michiel

2006-01-01

253

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.

Harms J.S.

1999-01-01

254

Potential transgenic routes to increase tree biomass.  

Science.gov (United States)

Biomass is a prime target for genetic engineering in forestry because increased biomass yield will benefit most downstream applications such as timber, fiber, pulp, paper, and bioenergy production. Transgenesis can increase biomass by improving resource acquisition and product utilization and by enhancing competitive ability for solar energy, water, and mineral nutrients. Transgenes that affect juvenility, winter dormancy, and flowering have been shown to influence biomass as well. Transgenic approaches have increased yield potential by mitigating the adverse effects of prevailing stress factors in the environment. Simultaneous introduction of multiple genes for resistance to various stress factors into trees may help forest trees cope with multiple or changing environments. We propose multi-trait engineering for tree crops, simultaneously deploying multiple independent genes to address a set of genetically uncorrelated traits that are important for crop improvement. This strategy increases the probability of unpredictable (synergistic or detrimental) interactions that may substantially affect the overall phenotype and its long-term performance. The very limited ability to predict the physiological processes that may be impacted by such a strategy requires vigilance and care during implementation. Hence, we recommend close monitoring of the resultant transgenic genotypes in multi-year, multi-location field trials. PMID:24094056

Dubouzet, Joseph G; Strabala, Timothy J; Wagner, Armin

2013-11-01

255

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

256

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

257

Rheumatic manifestations of inflammatory bowel disease  

Directory of Open Access Journals (Sweden)

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

258

Controlling transgene expression to study Xenopus laevis metamorphosis  

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Sperm-mediated transgenesis of Xenopus laevis is the first application of genetic methodology to an amphibian. However, some transgenes are lethal when they are expressed constitutively. To study the influence of these genes on amphibian metamorphosis and to generate F1 progeny from mature transgenic adults, these transgenes must be placed under the control of an inducible system so that they can be activated at specific times in development. We show that two well known binary inducible gene ...

Das, Biswajit; Brown, Donald D.

2004-01-01

259

Variable patterns of expression of luciferase in transgenic tobacco leaves.  

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A carboxyl-terminally modified firefly luciferase, encoded as a gene fusion to the neomycin phosphotransferase gene (which confers kanamycin resistance), was found to be enzymatically active for both enzymes when expressed in bacteria and in transgenic plants. A military-type starlight vision system was used to conveniently analyze the pattern of gene expression in transgenic tobacco plant leaves. Transgenic tobacco plants which expressed luciferase uniformly in all areas of the leaf, and ass...

1990-01-01

260

The Applications of Transgenic Rabbits in Agriculture and Biomedicine  

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During the last decades transgenic rabbits have provided suitable biological model for regulating and manipulating of interest genes. Several studies showed transgenic rabbits can be produced by different methods. In the last two decades pronucleotide microinjection was conventional and common method for produce transgenic rabbits but the current studies demonstrated that SMGT method provides a simple and straightforward technology to introduce DNA into rabbits which has many advantages in co...

Zabetian, M.; Tahmoorespur, M.; Kh. Hosseini

2011-01-01

 
 
 
 
261

Neurologic and motor dysfunctions in APP transgenic mice  

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The discovery of gene mutations underlying autosomal dominant Alzheimer’s disease has enabled researchers to reproduce several hallmarks of this disorder in transgenic mice, notably the formation of A? plaques in brain and cognitive deficits. APP transgenic mutants have also been investigated with respect to survival rates, neurologic functions, and motor coordination, which are all susceptible to alteration in Alzheimer dementia. Several transgenic lines expressing human mutated or wild-t...

Lalonde, Robert; Fukuchi, Ken-ichiro; Strazielle, Catherine

2012-01-01

262

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. The results indicate that an edi...

Yoshida, Taiji; Kimura, Eiichi; Koike, Setsuo; Nojima, Jun; Futai, Eugene; Sasagawa, Noboru; Watanabe, Yuichiro; Ishiura, Shoichi

2011-01-01

263

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

2005-05-06

264

Transgene elimination in genetically modified dry bean and soybean lines.  

Science.gov (United States)

Transgene elimination is a poorly studied phenomenon in plants. We made genetic and molecular studies of a transgenic dry bean line immune to bean golden mosaic geminivirus and a soybean line. In both lines, the transgenes were stable during the vegetative phase but were eliminated during meiosis. Due to its potential biotechnological value, this transgenic line was micropropagated by grafting and the vegetative copies were studied for more than two years. More than 300 plants of progeny were obtained during this period, demonstrating that the phenomenon of elimination was consistently repeated and offering an opportunity for detailed study of transgene elimination, including the characterization of the integration sites. Cloning and sequencing of the transgenic loci, reciprocal crosses to untransformed plants, genomic DNA blots, and GUS assays were performed in the transgenic lines. Based on the molecular and genetic characterization, possible mechanisms involved in transgene elimination include intrachromosomal recombination, genetic instability resulting from the tissue culture manipulations, and co-elimination of transgenes, triggered by a process of genome defense. PMID:16110439

Romano, Eduardo; Soares, Alexandre; Proite, Karina; Neiva, Suzana; Grossi, Maíra; Faria, Josias C; Rech, Elíbio L; Aragão, Francisco J L

2005-01-01

265

Development of a novel transgenic rice with hypocholesterolemic activity via high-level accumulation of the ?' subunit of soybean ?-conglycinin.  

Science.gov (United States)

Soybean 7S globulin, known as ?-conglycinin, has been shown to regulate human plasma cholesterol and triglyceride levels. Furthermore, the ?' subunit of ?-conglycinin has specifically been shown to possess low-density lipoprotein (LDL)-cholesterol-lowering activity. Therefore, accumulation of the ?' subunit of ?-conglycinin in rice seeds could lead to the production of new functional rice that could promote human health. Herein, we used the low-glutelin rice mutant 'Koshihikari' (var. a123) and suppressed its glutelins and prolamins, the major seed storage proteins of rice, by RNA interference. The accumulation levels of the ?' subunit in the lines with suppressed glutelin and prolamin levels were >20 mg in 1 g of rice seeds, which is considerably higher than those in previous studies. Oral administration of the transgenic rice containing the ?' subunit exhibited a hypocholesterolemic activity in rats; the serum total cholesterol and LDL cholesterol levels were significantly reduced when compared to those of the control rice (var. a123). The cholesterol-lowering action by transgenic rice accumulating the ?' subunit induces a significant increase in fecal bile acid excretion and a tendency to increase in fecal cholesterol excretion. This is the first report that transgenic rice exhibits a hypocholesterolemic activity in rats in vivo by using the ?-conglycinin ?' subunit. PMID:24676962

Cabanos, Cerrone; Kato, Naoki; Amari, Yoshiki; Fujiwara, Keigo; Ohno, Tomoki; Shimizu, Kousuke; Goto, Tsuyoshi; Shimada, Masaya; Kuroda, Masaharu; Masuda, Taro; Takaiwa, Fumio; Utsumi, Shigeru; Nagaoka, Satoshi; Maruyama, Nobuyuki

2014-08-01

266

Comparative characterization of mesenchymal stem cells from eGFP transgenic and non-transgenic mice  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Adipose derived- and bone marrow-derived murine mesenchymal stem cells (mMSCs may be used to study stem cell properties in an in vivo setting for the purposes of evaluating therapeutic strategies that may have clinical applications in the future. If these cells are to be used for transplantation, the question arises of how to track the administered cells. One solution to this problem is to transplant cells with an easily identifiable genetic marker such as enhanced green fluorescent protein (eGFP. This protein is fluorescent and therefore does not require a chemical substrate for identification and can be visualized in living cells. This study seeks to characterize and compare adipose derived- and bone marrow-derived stem cells from C57Bl/6 mice and eGFP transgenic C57Bl/6 mice. Results The expression of eGFP does not appear to affect the ability to differentiate along adipogenic or osteogenic lineages; however it appears that the tissue of origin can influence differentiation capabilities. The presence of eGFP had no effect on cell surface marker expression, and mMSCs derived from both bone marrow and adipose tissue had similar surface marker profiles. There were no significant differences between transgenic and non-transgenic mMSCs. Conclusion Murine adipose derived and bone marrow derived mesenchymal stem cells from non-transgenic and eGFP transgenic C57Bl/6 mice have very similar characterization profiles. The availability of mesenchymal stem cells stably expressing a genetic reporter has important applications for the advancement of stem cell research.

Bunnell Bruce A

2009-01-01

267

Transgenes sustain epigeal insect biodiversity in diversified vegetable farm systems.  

Science.gov (United States)

Many ecological studies have focused on the effects of transgenes in field crops, but few have considered multiple transgenes in diversified vegetable systems. We compared the epigeal, or soil surface-dwelling, communities of Coleoptera and Formicidae between transgenic and isoline vegetable systems consisting of sweet corn, potato, and acorn squash, with transgenic cultivars expressing Cry1(A)b, Cry3, or viral coat proteins. Vegetables were grown in replicated split plots over 2 yr with integrated pest management (IPM) standards defining insecticide use patterns. More than 77.6% of 11,925 insects from 1,512 pitfall traps were identified to species, and activity density was used to compare dominance distribution, species richness, and community composition. Measures of epigeal biodiversity were always equal in transgenic vegetables, which required fewer insecticide applications than their near isolines. There were no differences in species richness between transgenic and isoline treatments at the farm system and individual crop level. Dominance distributions were also similar between transgenic and isoline farming systems. Crop type, and not genotype, had a significant influence on Carabidae and Staphylinidae community composition in the first year, but there were no treatment effects in the second year, possibly because of homogenizing effects of crop rotations. Communities were more influenced by crop type, and possibly crop rotation, than by genotype. The heterogeneity of crops and rotations in diversified vegetable farms seems to aid in preserving epigeal biodiversity, which may be supplemented by reductions in insecticide use associated with transgenic cultivars. PMID:17349138

Leslie, T W; Hoheisel, G A; Biddinger, D J; Rohr, J R; Fleischer, S J

2007-02-01

268

Bacterial Diversity in Rhizospheres of Nontransgenic and Transgenic Corn  

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Bacterial diversity in transgenic and nontransgenic corn rhizospheres was determined. In greenhouse and field studies, metabolic profiling and molecular analysis of 16S rRNAs differentiated bacterial communities among soil textures but not between corn varieties. We conclude that bacteria in corn rhizospheres are affected more by soil texture than by cultivation of transgenic varieties.

Fang, Min; Kremer, Robert J.; Motavalli, Peter P.; Davis, Georgia

2005-01-01

269

Efficiency of human lactoferrin transgenic donor cell preparation for SCNT.  

Science.gov (United States)

The combination of somatic cell nuclear transfer (SCNT) and transgenic technology leads to the production of transgenic cloned animals, wherein the preparation of competent transgenic donor cells is the pivotal upstream step. The purpose of this study was to establish an efficient procedure to prepare human lactoferrin (hLTF) transgenic donor cells for SCNT. Thus, two cell culture systems were employed: caprine mammary epithelial cells (for evaluation of the hTLF transgenic expression in vitro), and fetal-derived fibroblast cells (for identification of competent transgenic donor cells). Induced by hormonal signals, recombinant hLTF was detected in the supernatant of transfected mammary epithelial cells by Western blot. Reliable hLTF transgenic fibroblast cell clones were identified by screening with multiple PCR amplification, EGFP fluorescence, and chromosomal counting (32.5+/-2.3%). This study may provide an effective upstream system to prepare SCNT donor cells for the production of human recombinant pharmaceuticals from the milk of transgenic animals. PMID:18804853

Zhao, M T; Lin, H; Liu, F J; Quan, F S; Wang, G H; Liu, J; Hua, S; Zhang, Y

2009-01-15

270

Effects of TGF-?1 and VEGF-A transgenes on the osteogenic potential of bone marrow stromal cells in vitro and in vivo.  

Science.gov (United States)

An exogenous supply of growth factors and bioreplaceable scaffolds may help bone regeneration. The aim of this study was to examine the effects of TGF-?1 and VEGF-A transgenes on the osteogenic potential of bone marrow stromal cells. Rat bone marrow stromal cells were transfected with plasmids encoding mouse TGF-?1 and/or VEGF-A complementary DNAs and cultured for up to 28 days. Furthermore, collagen scaffolds carrying combinations of the plasmids-transfected cells were implanted subcutaneously in rats. The transgenes increased alkaline phosphatase activity, enhanced mineralized nodule formation, and elevated osteogenic gene expressions in vitro. In vivo, messenger RNA expression of osteogenic genes such as BMPs and Runx2 elevated higher by the transgenes. The data indicate that exogenous TGF-?1 and VEGF-A acted synergistically and could induce osteoblastic differentiation of bone marrow stromal cells in both cell culture and an animal model. The results may provide valuable information to optimize protocols for transgene-and-cell-based tissue engineering. PMID:22962632

Kuroda, Shinji; Sumner, Dale R; Virdi, Amarjit S

2012-01-01

271

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

272

Zebrafish transgenic Enhancer TRAP line database (ZETRAP  

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Full Text Available 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 been collected and compiled in the form of an online database – the Zebrafish Enhancer TRAP lines database (ZETRAP. Conclusion ZETRAP is a web-based system that provides data and information to the scientific community about the developmental, genetic and genomic aspects of transgenic zebrafish lines obtained using Tol2 transposon-mediated transgenesis. The current version (version 1.0 contains description of 27 ET lines that express EGFP in various organs and tissues, for example, heart, brain, notochord, gut, etc. It also includes information on insertion sites of the Tol2 transposon in these lines.

Emelyanov Alexander

2006-02-01

273

Transgenic crops. Processes, products and problems  

International Nuclear Information System (INIS)

Transgenic crops are a natural extension of plant breeding technologies, offering new opportunities for increasing the productivity of agriculture and reducing the cost of food production, for increasing the appeal, nutritional content and quality of fresh and processed foods, and for reducing the environmental damage of agricultural practices. These new transgenic traits will be combined with continuing improvements in the latest varieties developed via breeding technologies, spurring investment in both. These technologies are inherently compatible with and necessary for meeting the challenges now facing the world, namely, economic growth and development, environmental protection and remediation, and human needs for food, shelter and a decent quality of life. The first products from genetically engineered crops are beginning to enter commerce. This is a critical time for issues that will shape public acceptance and for adoption of regulatory and trade policies that encourage rather than discourage investment in and use of this technology. Further investment in the tools for transforming crops and in the basic and applied sciences that will provide a pipeline of new genes is also needed. (author). 24 refs, 1 tab

1995-11-01

274

Accumulation of fructose polymers in transgenic tobacco.  

Science.gov (United States)

Fructan, a polyfructose molecule, is a storage compound in a limited number of plant species. Usually these species accumulate fructan with a low degree of polymerization (DP) and most of these plants have properties which preclude their use as a fructan source. With the eventual aim of allowing the accumulation of high DP fructans in non-fructan storing plants, we have investigated whether carbohydrate flow in the plant cell can be directed to produce this polymer. For this purpose the SacB gene from Bacillus subtilis, which encodes levansucrase, was modified and introduced into tobacco plants. Transgenic plants containing the sacB gene accumulate fructans. The size and properties of this fructan are similar to fructan produced by Bacillus subtilis, and is stable in plants. Although the level of fructan accumulation in the transgenic tobacco plants ranged from 3-8 percent of the dry weight, no levansucrase mRNA or protein could be detected in these plants. Extension of this work should permit the production of this high molecular weight biopolymer in crop plants for applications in food and non-food products. PMID:7764488

Ebskamp, M J; van der Meer, I M; Spronk, B A; Weisbeek, P J; Smeekens, S C

1994-03-01

275

A transgenic mouse model for lung adenocarcinoma.  

Science.gov (United States)

Lung cancer is a leading cause of tumor-related deaths in humans but its origin and development are poorly understood. To study the biology of these tumors, appropriate animal and cell culture models will be of eminent importance. Uteroglobin is a marker protein for the nonciliated epithelial Clara cells lining the respiratory and terminal bronchioli of the lung. We have used the promoter and 5'-flanking sequences of the rabbit uteroglobin gene to target expression of the SV40 T antigen to the lung of transgenic mice. All transgenic founders as well as the descendants from an established line, UT7.1, developed multifocal bronchioloalveolar adenocarcinomas originating from Clara cells. At least three different stages in tumor development with progressive loss of the differentiated phenotype can be distinguished by immunohistochemical data and in situ hybridization. Only in the initial stage did bronchiolar cells express both uteroglobin and SV40 T antigen, whereas at later stages, only SV40 T antigen was detected, and the most advanced tumors were negative for both proteins. Starting from the lungs of UT7.1 mice, a bronchiolar cell line was established that maintains the features of differentiated Clara cells. This system provides a useful model for further studying the development and progression of lung adenocarcinomas in vivo and in vitro. PMID:7718490

Sandmöller, A; Halter, R; Suske, G; Paul, D; Beato, M

1995-01-01

276

Spatial and temporal control of transgene expression in zebrafish.  

Science.gov (United States)

Transgenic zebrafish research has provided valuable insights into gene functions and cell behaviors directing vertebrate development, physiology, and disease models. Most approaches use constitutive transgene expression and therefore do not provide control over the timing or levels of transgene induction. We describe an inducible gene expression system that uses new tissue-specific zebrafish transgenic lines that express the Gal4 transcription factor fused to the estrogen-binding domain of the human estrogen receptor. We show these Gal4-ERT driver lines confer rapid, tissue-specific induction of UAS-controlled transgenes following tamoxifen exposure in both embryos and adult fish. We demonstrate how this technology can be used to define developmental windows of gene function by spatiotemporal-controlled expression of constitutively active Notch1 in embryos. Given the array of existing UAS lines, the modular nature of this system will enable many previously intractable zebrafish experiments. PMID:24643048

Akerberg, Alexander A; Stewart, Scott; Stankunas, Kryn

2014-01-01

277

Pituitary-directed leukemia inhibitory factor transgene forms Rathke's cleft cysts and impairs adult pituitary function. A model for human pituitary Rathke's cysts.  

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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

278

Effects of TGF-?1 and VEGF-A transgenes on the osteogenic potential of bone marrow stromal cells in vitro and in vivo  

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An exogenous supply of growth factors and bioreplaceable scaffolds may help bone regeneration. The aim of this study was to examine the effects of TGF-?1 and VEGF-A transgenes on the osteogenic potential of bone marrow stromal cells. Rat bone marrow stromal cells were transfected with plasmids encoding mouse TGF-?1 and/or VEGF-A complementary DNAs and cultured for up to 28 days. Furthermore, collagen scaffolds carrying combinations of the plasmids-transfected cells were implanted subcutaneo...

Kuroda, Shinji; Sumner, Dale R.; Virdi, Amarjit S.

2012-01-01

279

Inheritance of transgenes in transgenic Bt lines resistance to Helicoerpa armigera in upland cotton.  

Science.gov (United States)

Six transgenic Bt cotton cultivars (lines) including GKsu12, GK19, MR1, GK5, 109B, and SGK1 are highly resistant to bollworm from the seedling to boll-setting stages in bioassays with detached cotton leaves, though there are differences in resistant level and Bt toxin content in these transgenic cottons. Genetics analysis reveals that the resistance to Helicoverpa armigera in these six transgenic Bt cotton cultivars (lines) are controlled by one pair of dominant genes. Allelic tests further demonstrate some populations are in Mendel segregation for two nonallelic genes, i.e., the inserted Bt gene in GKsu12 is nonallelic to that of SGK1, GK5, 109B, and GK19 and Bt genes in GK19 and SGK1 are likely inserted in the same or in close proximity (genetically closely linked), while some F(2) produce abnormal segregation patterns, with a segregation of resistance to Helicoerpa armigera vary between 15:1 and 3:1, though their Bt segregation fit into 15:1 by PCR analysis, suggesting Bt gene silence in these populations. Two genes silence may occur in these populations due to the homologous sequence by crossing since the silenced individuals accounted for 1/16 of the F(2) populations for allelic test. To those silenced populations, one of their parents all showed high resistance to bollworm. PMID:23143492

Zhang, Baolong; Guo, Wangzhen; Zhang, Tianzhen

2013-01-01

280

Enhancement of plasmid-mediated transgene expression.  

Science.gov (United States)

A large number of studies aimed at the treatment of cancer, autoimmune and metabolic diseases, neurodegenerative disorders, allergic diseases, as well as muscle disorders strengthen the fact that gene therapy could represent an alternative method to treat human diseases where conventional approaches are less effective. To improve transgene expression from plasmid vectors, DNA nuclear targeting sequences (DTSs) can be introduced in a vector backbone to increase in vivo expression up to 20-fold using electroporation (EP) delivery in muscle tissue. The purpose of this chapter is to represent a step-by-step strategy for the construction of a plasmid vector with enhanced efficiency of nuclear plasmid uptake and the methodic for the in vivo efficiency evaluation of the obtained expression vector. PMID:24715279

Fioretti, Daniela; Iurescia, Sandra; Rinaldi, Monica

2014-01-01

 
 
 
 
281

Magnetic biomineralisation in Huntington's disease transgenic mice  

International Nuclear Information System (INIS)

The concentration levels of biogenic magnetite nanoparticles in transgenic R6/2 Huntington's disease (HD) mice have been investigated, using seven control and seven HD mice each from an 8 week-old litter and from a 12 week-old litter. Hysteresis and isothermal remnant magnetisation data were collected on a SQUID magnetometer, and analysed using a model comprising dia/paramagnetic, ferrimagnetic and superparamagnetic contributions, to extract the magnetite and ferritin concentrations present. It was found that magnetite was present in both superparamagnetic and blocked states. A larger spread and higher concentration of magnetite levels was found in the diseased mice for both the 8 week-old and 12 week-old batches, compared to the controls

2005-01-01

282

Gene therapy: X-SCID transgene leukaemogenicity.  

Science.gov (United States)

Gene therapy has been remarkably effective for the immunological reconstitution of patients with severe combined immune deficiency, but the occurrence of leukaemia in a few patients has stimulated debate about the safety of the procedure and the mechanisms of leukaemogenesis. Woods et al. forced high expression of the corrective therapeutic gene IL2RG, which encodes the gamma-chain of the interleukin-2 receptor, in a mouse model of the disease and found that tumours appeared in a proportion of cases. Here we show that transgenic IL2RG does not necessarily have potent intrinsic oncogenic properties, and argue that the interpretation of this observation with respect to human trials is overstated. PMID:16988659

Thrasher, Adrian J; Gaspar, H Bobby; Baum, Christopher; Modlich, Ute; Schambach, Axel; Candotti, Fabio; Otsu, Makoto; Sorrentino, Brian; Scobie, Linda; Cameron, Ewan; Blyth, Karen; Neil, Jim; Abina, Salima Hacein-Bey; Cavazzana-Calvo, Marina; Fischer, Alain

2006-09-21

283

Magnetic biomineralisation in Huntington's disease transgenic mice  

Science.gov (United States)

The concentration levels of biogenic magnetite nanoparticles in transgenic R6/2 Huntington's disease (HD) mice have been investigated, using seven control and seven HD mice each from an 8 week-old litter and from a 12 week-old litter. Hysteresis and isothermal remnant magnetisation data were collected on a SQUID magnetometer, and analysed using a model comprising dia/paramagnetic, ferrimagnetic and superparamagnetic contributions, to extract the magnetite and ferritin concentrations present. It was found that magnetite was present in both superparamagnetic and blocked states. A larger spread and higher concentration of magnetite levels was found in the diseased mice for both the 8 week-old and 12 week-old batches, compared to the controls.

Beyhum, W.; Hautot, D.; Dobson, J.; Pankhurst, Q. A.

2005-01-01

284

High-value products from transgenic maize.  

Science.gov (United States)

Maize (also known as corn) is a domesticated cereal grain that has been grown as food and animal feed for tens of thousands of years. It is currently the most widely grown crop in the world, and is used not only for food/feed but also to produce ethanol, industrial starches and oils. Maize is now at the beginning of a new agricultural revolution, where the grains are used as factories to synthesize high-value molecules. In this article we look at the diversity of high-value products from maize, recent technological advances in the field and the emerging regulatory framework that governs how transgenic maize plants and their products are grown, used and traded. PMID:20816943

Naqvi, Shaista; Ramessar, Koreen; Farré, Gemma; Sabalza, Maite; Miralpeix, Bruna; Twyman, Richard M; Capell, Teresa; Zhu, Changfu; Christou, Paul

2011-01-01

285

Direct Renin Inhibition Improves Systemic Insulin Resistance and Skeletal Muscle Glucose Transport in a Transgenic Rodent Model of Tissue Renin Overexpression  

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Renin is the rate-limiting enzyme in renin-angiotensin system (RAS) activation. We sought to determine the impact of renin inhibition on whole-body insulin sensitivity and skeletal muscle RAS, oxidative stress, insulin signaling, and glucose transport in the transgenic TG(mRen2)27 rat (Ren2), which manifests increased tissue RAS activity, elevated serum aldosterone, hypertension, and insulin resistance. Young (aged 6–9 wk) Ren2 and age-matched Sprague Dawley control rats were treated with a...

Lastra, Guido; Habibi, Javad; Whaley-connell, Adam T.; Manrique, Camila; Hayden, Melvin R.; Rehmer, Jenna; Patel, Kamlesh; Ferrario, Carlos; Sowers, James R.

2009-01-01

286

Generation and characterization of human heme oxygenase-1 transgenic pigs.  

Science.gov (United States)

Xenotransplantation using transgenic pigs as an organ source is a promising strategy to overcome shortage of human organ for transplantation. Various genetic modifications have been tried to ameliorate xenograft rejection. In the present study we assessed effect of transgenic expression of human heme oxygenase-1 (hHO-1), an inducible protein capable of cytoprotection by scavenging reactive oxygen species and preventing apoptosis caused by cellular stress during inflammatory processes, in neonatal porcine islet-like cluster cells (NPCCs). Transduction of NPCCs with adenovirus containing hHO-1 gene significantly reduced apoptosis compared with the GFP-expressing adenovirus control after treatment with either hydrogen peroxide or hTNF-? and cycloheximide. These protective effects were diminished by co-treatment of hHO-1 antagonist, Zinc protoporphyrin IX. We also generated transgenic pigs expressing hHO-1 and analyzed expression and function of the transgene. Human HO-1 was expressed in most tissues, including the heart, kidney, lung, pancreas, spleen and skin, however, expression levels and patterns of the hHO-1 gene are not consistent in each organ. We isolate fibroblast from transgenic pigs to analyze protective effect of the hHO-1. As expected, fibroblasts derived from the hHO-1 transgenic pigs were significantly resistant to both hydrogen peroxide damage and hTNF-? and cycloheximide-mediated apoptosis when compared with wild-type fibroblasts. Furthermore, induction of RANTES in response to hTNF-? or LPS was significantly decreased in fibroblasts obtained from the hHO-1 transgenic pigs. These findings suggest that transgenic expression of hHO-1 can protect xenografts when exposed to oxidative stresses, especially from ischemia/reperfusion injury, and/or acute rejection mediated by cytokines. Accordingly, hHO-1 could be an important candidate molecule in a multi-transgenic pig strategy for xenotransplantation. PMID:23071605

Yeom, Hye-Jung; Koo, Ok Jae; Yang, Jaeseok; Cho, Bumrae; Hwang, Jong-Ik; Park, Sol Ji; Hurh, Sunghoon; Kim, Hwajung; Lee, Eun Mi; Ro, Han; Kang, Jung Taek; Kim, Su Jin; Won, Jae-Kyung; O'Connell, Philip J; Kim, Hyunil; Surh, Charles D; Lee, Byeong-Chun; Ahn, Curie

2012-01-01

287

[Effects of phytase transgenic corn planting on soil nematode community].  

Science.gov (United States)

A healthy soil ecosystem is essential for nutrient cycling and energy conversion, and the impact of exogenous genes from genetically modified crops had aroused wide concerns. Phytase transgenic corn (i. e., the inbred line BVLA430101) was issued a bio-safety certificate on 27 September 2009 in China, which could improve the efficiency of feed utilization, reduce environmental pollution caused by animal manure. In this study, the abundance of trophic groups, community structure and ecological indices of soil nematodes were studied over the growing cycle of phytase transgenic corn (ab. transgenic corn) and control conventional parental corn (ab. control corn) in the field. Totally 29 and 26 nematode genera were isolated from transgenic corn and control corn fields, respectively. The abundances of bacterivores and omnivores-predators, the total number of soil nematodes, and the Shannon index (H) were significantly greater under transgenic corn than under control corn, while the opposite trend was found for the relative abundance of herbivores and the maturity index (Sigma MI) of soil nematodes. Repeated-measures analysis of variance (ANOVA) did not detect any significant effects of transgenic corn on the composition and abundance of nematode trophic groups and ecological indices of soil nematodes. Furthermore, the Student-T test showed that the abundances of bacterivores and omnivores-predators and the total number of soil nematodes during the milk-ripe stage were significant higher in the transgenic corn field than in the control corn field. The effects of transgenic corn planting on soil nematodes might be related to the increase in the nitrogen content of field soil under transgenic corn compared to control corn. PMID:25011306

Zhao, Zong-Chao; Su, Ying; Mou, Wen-Ya; Liu, Man-Qiang; Chen, Xiao-Yun; Chen, Fa-Jun

2014-04-01

288

Generation and Characterization of Human Heme Oxygenase-1 Transgenic Pigs  

Science.gov (United States)

Xenotransplantation using transgenic pigs as an organ source is a promising strategy to overcome shortage of human organ for transplantation. Various genetic modifications have been tried to ameliorate xenograft rejection. In the present study we assessed effect of transgenic expression of human heme oxygenase-1 (hHO-1), an inducible protein capable of cytoprotection by scavenging reactive oxygen species and preventing apoptosis caused by cellular stress during inflammatory processes, in neonatal porcine islet-like cluster cells (NPCCs). Transduction of NPCCs with adenovirus containing hHO-1 gene significantly reduced apoptosis compared with the GFP-expressing adenovirus control after treatment with either hydrogen peroxide or hTNF-? and cycloheximide. These protective effects were diminished by co-treatment of hHO-1 antagonist, Zinc protoporphyrin IX. We also generated transgenic pigs expressing hHO-1 and analyzed expression and function of the transgene. Human HO-1 was expressed in most tissues, including the heart, kidney, lung, pancreas, spleen and skin, however, expression levels and patterns of the hHO-1 gene are not consistent in each organ. We isolate fibroblast from transgenic pigs to analyze protective effect of the hHO-1. As expected, fibroblasts derived from the hHO-1 transgenic pigs were significantly resistant to both hydrogen peroxide damage and hTNF-? and cycloheximide-mediated apoptosis when compared with wild-type fibroblasts. Furthermore, induction of RANTES in response to hTNF-? or LPS was significantly decreased in fibroblasts obtained from the hHO-1 transgenic pigs. These findings suggest that transgenic expression of hHO-1 can protect xenografts when exposed to oxidative stresses, especially from ischemia/reperfusion injury, and/or acute rejection mediated by cytokines. Accordingly, hHO-1 could be an important candidate molecule in a multi-transgenic pig strategy for xenotransplantation.

Yang, Jaeseok; Cho, Bumrae; Hwang, Jong-Ik; Park, Sol Ji; Hurh, Sunghoon; Kim, Hwajung; Lee, Eun Mi; Ro, Han; Kang, Jung Taek; Kim, Su Jin; Won, Jae-Kyung; O'Connell, Philip J.; Kim, Hyunil; Surh, Charles D.; Lee, Byeong-Chun; Ahn, Curie

2012-01-01

289

Ectopic growth of hippocampal mossy fibers in a mutated GAP-43 transgenic mouse with impaired spatial memory retention  

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In a previous study, it was shown that transgenic mice, designated G-NonP, forget the location of a water maze hidden platform when tested 7 days after the last training day (Holahan and Routtenberg, 2008). The memory loss in G-NonP mice might be related to altered hippocampal architecture suggested by the fact that in the rat, 7 days after water maze training, there is discernible mossy fiber (MF) growth (Holahan et al., 2006; Rekart et al., 2007). In the present report, we studied the distr...

Holahan, Matthew R.; Honegger, Kyle S.; Routtenberg, Aryeh

2010-01-01

290

Increased Expression of the Na,K-ATPase alpha4 Isoform Enhances Sperm Motility in Transgenic Mice1  

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The Na,K-ATPase alpha4 (ATP1A4) isoform is specifically expressed in male germ cells and is highly prevalent in spermatozoa. Although selective inhibition of alpha4 activity with ouabain has been shown to affect sperm motility, a more direct analysis of the role of this isoform in sperm movement has not yet been demonstrated. To establish this, we engineered transgenic mice that express the rat alpha4 isoform fused to green fluorescent protein in male germ cells, under the control of the mous...

Jimenez, Tamara; Sanchez, Gladis; Mcdermott, Jeffrey P.; Nguyen, Anh-nguyet; Kumar, T. Rajendra; Blanco, Gustavo

2011-01-01

291

Generating transgenic frog embryos by restriction enzyme mediated integration (REMI).  

Science.gov (United States)

Here we present a protocol for generating transgenic embryos in Xenopus laevis and Xenopus tropicalis. The method includes three steps: (1) The preparation of high-speed egg extracts, which facilitates the replacement of protamines in sperm nuclei with nucleosomes and decondenses the chromatin of sperm nuclei; (2) The isolation of sperm nuclei; and (3) The mixing of sperm nuclei, restriction enzyme, and high-speed extract in vitro, following by nuclear transplantation into unfertilized eggs to generate the transgenic embryos. This procedure generates non-mosaic transgenic embryos at high frequency and efficiency. PMID:22956089

Ishibashi, Shoko; Kroll, Kristen L; Amaya, Enrique

2012-01-01

292

Transgenic rice expressing amyloid ?-peptide for oral immunization.  

Science.gov (United States)

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. The results indicate that an edible vaccine against AD using rice may be feasible. A vaccine derived from rice would be far cheaper than existing medical vaccines. PMID:21448341

Yoshida, Taiji; Kimura, Eiichi; Koike, Setsuo; Nojima, Jun; Futai, Eugene; Sasagawa, Noboru; Watanabe, Yuichiro; Ishiura, Shoichi

2011-01-01

293

Regeneration of transgenic citrus plants under non selective conditions results in high-frequency recovery of plants with silenced transgenes.  

Science.gov (United States)

Insertion of foreign DNA into plant genomes frequently results in the recovery of transgenic plants with silenced transgenes. To investigate to what extent regeneration under selective conditions limits the recovery of transgenic plants showing gene silencing in woody species, Mexican lime [ Citrus aurantifolia (Christm.) Swing.] plants were transformed with the p25 coat protein gene of Citrus tristeza virus (CTV) with or without selection for nptII and uidA. Strikingly, more than 30% of the transgenic limes regenerated under non-selective conditions had silenced transgenes, and in all cases silencing affected all the three transgenes incorporated. These results indicate that the frequency of transgene silencing may be greatly underestimated when the rate of silencing is estimated from the number of regenerants obtained under selective conditions. To our knowledge, this is the first report in which the frequency of gene silencing after transformation has been quantified. When the integration pattern of T-DNA was analyzed in silenced and non-silenced lines, it was observed that inverted repeats as well as direct repeats and even single integrations were able to trigger gene silencing. Gene silencing has often been associated with the insertion of DNA sequences as inverted repeats. Interestingly, here, direct repeats and single-copy insertions were found in both silenced and non-silenced lines, suggesting that the presence of inverted-repeat T-DNAs and the subsequent formation of dsRNAs triggering gene silencing cannot account for all silencing events. PMID:12111562

Domínguez, A; Fagoaga, C; Navarro, L; Moreno, P; Peña, L

2002-06-01

294

Transgenic approaches to a non-transgenic release of sterile male Lepidoptera  

International Nuclear Information System (INIS)

Successful implementation of the Sterile Insect Technique (SIT) against codling moth, Cydia pomonella (L.) (Tortricidae), in British Columbia, Canada, resulted in demands for the expansion of codling moth SIT and a related suppression strategy, radiation-induced inherited sterility (IS), in other countries. In the current SIT programme, both sterile males and females are released to control the pest population. There are compelling reasons to believe that both codling moth SIT and IS would benefit if efficient ways could be found to produce and release only males. Recently, a new scheme for genetic sexing in Lepidoptera has been proposed. The scheme is based on the construction of transgenic females carrying a dominant conditional lethal gene in the female-determining chromosome W. Following this scheme we intend to develop transgenic sexing strains in the codling moth. This requires basic knowledge of codling moth genome and appropriate molecular tools for codling moth transgenesis. We performed a detailed analysis of codling moth karyotype with a particular focus on the identification and characterization of sex chromosomes. Here we summarize our data on codling moth cytogenetics and discuss the potential of codling moth sex chromosomes for their use in developing transgenic sexing strains. The karyotype of codling moth consists of 2n=56 chromosomes, which can be classified into five groups according to their sizes: extra large (3 pairs), large (3 pairs), medium (15 pairs), small (5 pairs), and dot-like (2 pairs). Females are heterogametic with a W-Z sex chromosome pair, males are homogametic with two Z chromosomes. The W and Z chromosomes represent the two largest elements in female chromosome complements. While the Z is composed of euchromatin and resembles to autosomes, the W consists largely of heterochromatin. For successful development of transgenic sexing strains in the codling moth, it is required to insert a conditional dominant lethal mutation (a transgene) into the W chromosome. Theoretically, the transgene insertion is a matter of probability, which is dependent on the size of the W relative to the rest of the genome. In the codling moth, the W is one of two largest chromosomes, comprising about 4% of the female genome, which should make it a good target for transgenesis with the probability of insertion of 1 in 25 (if only females are included) or with the overall probability of 1 in 50 (since both female and male embryos are exposed). However, since the W chromosome is mainly composed of heterochromatin, silencing of the transgene expression might be a serious problem. Different ways how to overcome this problem are discussed. For further characterisation of the codling moth W chromosome we employed advanced methods of molecular cytogenetics, genomic in situ hybridisation (GISH) and comparative genomic hybridisation (CGH). GISH detected the W chromosome by strong binding of the Cy3-labelled, female-derived DNA probe. With CGH, both the Cy3-labelled female-derived probe and Fluor-X labelled male-derived probe evenly bound to the W. This suggested that the W is composed predominantly of repetitive DNA sequences occurring scattered in other chromosomes but accumulated in the W. Finally, we prepared W-specific probes by laser microdissection of the W chromatin followed by DOP-PCR and PCR labelling. The probes stained the W with a high specificity in a chromosome-painting manner. DNA fragments of the microdissected W chromatin were cloned and sequenced. The W-sequence analysis revealed no homology to any DNA sequenced so far. Several cloned sequences were found to originate exclusively from the W chromosome. These unique sequences can be very useful as molecular markers of the W chromosome in codling moth transgenesis. The demonstrated ways of W chromosome identification will facilitate the development of genetic sexing strains in the codling moth

2005-05-09

295

Rskalpha-actin/hIGF-1 transgenic mice with increased IGF-I in skeletal muscle and blood: impact on regeneration, denervation and muscular dystrophy.  

Science.gov (United States)

Human IGF-I was over-expressed in skeletal muscles of C57/BL6xCBA mice under the control of the rat skeletal alpha-actin gene promoter. RT-PCR verified expression of the transgene in skeletal muscle but not in the liver of 1- and 21-day old heterozygote transgenic mice. The concentration of endogenous mouse IGF-I, measured by an immunoassay which does not detect human IGF-I, was not significantly different between transgenic mice and wild-type littermates (9.5 +/- 0.8 and 13.3 +/- 1.9 ng/g in muscle; 158.3 +/- 18.6 and 132.9 +/- 33.1 ng/ml in plasma, respectively). In contrast, quantitation with antibodies to human IGF-I showed an increase in IGF-I of about 100 ng/ml in plasma and 150 ng/g in muscle of transgenic mice at 6 months of age. Transgenic males, compared to their age matched wild-type littermates, had a significantly higher body weight (38.6 +/- 0.53 g vs. 35.8 +/- 0.64 g at 6 months of age; P myofibrillar protein mass (1.62 +/- 0.045 vs. 1.49 +/- 0.048 g; P hypertrophy and no change in the proportion of slow type I myofibres in the limb muscles of Rskalpha-actin/hIGF-I transgenic mice at 3 or 6 months of age. Phenotypic changes in Rskalpha-actin/hIGF-I mice are likely to be due to systemic as well as autocrine/paracrine effects of overproduction of IGF-I due to expression of the human IGF-I transgene. The effect of muscle specific over-expression of Rskalpha-actin/hIGF-I transgene was tested on: (i) muscle regeneration in auto-transplanted whole muscle grafts; (ii) myofibre atrophy following sciatic nerve transection; and (iii) sarolemmal damage and myofibre necrosis in dystrophic mdx muscle. No beneficial effect of muscle specific over-expression of Rskalpha-actin/hIGF-I transgene was seen in these three experimental models. PMID:16716629

Shavlakadze, T; Boswell, J M; Burt, D W; Asante, E A; Tomas, F M; Davies, M J; White, J D; Grounds, M D; Goddard, C

2006-06-01

296

Transgenic potato plants with overexpression of dihydroflavonol reductase can serve as efficient nutrition sources.  

Science.gov (United States)

Potato (Solanum tuberosum) is considered to be one of the most important crops cultivated in Europe and the entire world. The tubers of the potato are characterized by rich starch and protein contents and high concentrations of antioxidants, such as vitamin C and flavonoids. Notably, the presence of the phenolic antioxidants is of high importance as they have health-related properties. They are known to reduce the incidence of atherosclerosis, prevent certain kinds of cancer, and aid with many other kinds of diseases. The aim of this study was to find the most efficient way to increase the content of phenolic antioxidants in potato tubers through transgenesis. The results showed that the most efficacious way to achieve this goal was the overexpression of the dihydroflavonol reductase gene (DFR). The produced transgenic potato plants served as a nutrition source for laboratory rats; the study has confirmed their nontoxicity and nutritional benefits on the tested animals. PMID:23692339

Kostyn, Kamil; Szatkowski, Michal; Kulma, Anna; Kosieradzka, Iwona; Szopa, Jan

2013-07-10

297

Comparison of semen characteristics and histological structure of the testis from transgenic and non-transgenic rabbits  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The aim of this study was to compare semen characteristics including sperm quantity, quality, and abnormalities, as well as histological structure of the testis of three-year old transgenic (human clotting factor, hFVIII, gene) and nontransgenic rabbits. For the experiment, 10 transgenic rabbits of F2 and F3 generations and 10 randomly selected non-transgenic males of the same breed and age were used as controls. All males were housed in individual cages, under a the same environmental condit...

Lukac, N.; Massanyi, P.; Flesarova, S.; Danko, J.; Makarevich, A. V.; Chrenek, P.

2009-01-01

298

Transgenic mice carrying a human mutant superoxide dismutase transgene develop neuronal cytoskeletal pathology resembling human amyotrophic lateral sclerosis lesions.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Mutations in the human Cu,Zn superoxide dismutase gene (SOD1) are found in 20% of kindreds with familial amyotrophic lateral sclerosis. Transgenic mice (line G1H) expressing a human SOD1 containing a mutation of Gly-93 --> Ala (G93A) develop a motor neuron disease similar to familial amyotrophic lateral sclerosis, but transgenic mice (line N1029) expressing a wild-type human SOD1 transgene do not. Because neurofilament (NF)-rich inclusions in spinal motor neurons are characteristic of amyotro...

Tu, P. H.; Raju, P.; Robinson, K. A.; Gurney, M. E.; Trojanowski, J. Q.; Lee, V. M.

1996-01-01

299

Sensitivity of a real-time PCR method for the detection of transgenes in a mixture of transgenic and non-transgenic seeds of papaya (Carica papaya L.)  

Science.gov (United States)

Background Genetically engineered (GE) ringspot virus-resistant papaya cultivars ‘Rainbow’ and ‘SunUp’ have been grown in Hawai’i for over 10 years. In Hawai’i, the introduction of GE papayas into regions where non-GE cultivars are grown and where feral non-GE papayas exist have been accompanied with concerns associated with transgene flow. Of particular concern is the possibility of transgenic seeds being found in non-GE papaya fruits via cross-pollination. Development of high-throughput methods to reliably detect the adventitious presence of such transgenic material would benefit both the scientific and regulatory communities. Results We assessed the accuracy of using conventional qualitative polymerase chain reaction (PCR) as well as real-time PCR-based assays to quantify the presence of transgenic DNA from bulk samples of non-GE papaya seeds. In this study, an optimized method of extracting high quality DNA from dry seeds of papaya was standardized. A reliable, sensitive real-time PCR method for detecting and quantifying viral coat protein (cp) transgenes in bulk seed samples utilizing the endogenous papain gene is presented. Quantification range was from 0.01 to 100 ng/?l of GE-papaya DNA template with a detection limit as low as 0.01% (10 pg). To test this system, we simulated transgene flow using known quantities of GE and non-GE DNA and determined that 0.038% (38 pg) GE papaya DNA could be detected using real-time PCR. We also validated this system by extracting DNA from known ratios of GE seeds to non-GE seeds of papaya followed by real-time PCR detection and observed a reliable detection limit of 0.4%. Conclusions This method for the quick and sensitive detection of transgenes in bulked papaya seed lots using conventional as well as real-time PCR-based methods will benefit numerous stakeholders. In particular, this method could be utilized to screen selected fruits from maternal non-GE papaya trees in Hawai’i for the presence of transgenic seed at typical regulatory threshold levels. Incorporation of subtle differences in primers and probes for variations in cp worldwide should allow this method to be utilized elsewhere when and if deregulation of transgenic papaya occurs.

2013-01-01

300

Transgenic mice expressing the human inducible Hsp70 have hippocampal neurons resistant to ischemic injury  

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Using transgenic mice constitutively expressing the human inducible Hsp70, we examined the role of Hsp70 on cell survival after focal cerebral oschemia. Twenty-four hours after premanent occlusion of the middle cerebral artery, no difference in infarct area was detected between Hsp70-transgenic and non-transgenic mice. In the non-transgenic mice, many pyramidal neurons of the ipsilateral hippocampus were observed to be pykontic. However, in all Hsp70-transgenic mice, hippocampal pyramidal neu...

Plumier, J. -c L.; Krueger, A. M.; Currie, R. W.; Kontoyiannis, D.; Kollias, G.; Pagoulatos, G. N.

1997-01-01

 
 
 
 
301

Effects of Transgenic Rice on Life History Traits of Daphnia magna in Life Table Experiments  

Digital Repository Infrastructure Vision for European Research (DRIVER)

To investigate the impacts of transgenic rice on freshwater organisms, we conducted two life tableexperiments using Daphnia magna for fifteen and twenty days, respectively. We examined life history traits suchas population growth rates (r), reproductive rates (R0), generation times, and survivorship. In the first experiment,we used non-drought-stressed transgenic and non-transgenic rice harvested in 2005. In the second study, weused non-transgenic and transgenic rice harvested in 2006 followi...

2007-01-01

302

The transgenic mouse assay as an alternative test method for regulatory carcinogenicity studies--implications for REACH.  

Science.gov (United States)

REACH, an EU regulation that requires the submission of safety data in support of the protection of human and environmental health, mandates that registration should be achieved with the minimum amount of animal testing possible. Under REACH, a two-year carcinogenicity assay may be required for certain chemicals produced at >1000 metric tonnes per year. In addition, some chemicals that are found to be genotoxic will also require testing. Alternative methods have been explored in an attempt to improve the predictivity of this bioassay as well as to reduce the number of animals used for such testing. This research has focused on the use of transgenic/knockout mouse models. Study results from selected models indicate that they are useful in hazard identification, even if they are not entirely suitable for risk assessment on their own. Carcinogenic hazard assessment can be greatly enhanced and animal use reduced if the traditional two-year rat bioassay is combined with a well conducted transgenic mouse assay. Importantly, the use of transgenic animals to supplement a traditional two-year carcinogenicity study may help reduce the number of false negatives, one of the unstated goals of REACH via the precautionary principle. PMID:19126422

Wells, Monique Y; Williams, E Spencer

2009-03-01

303

Polycythemia in transgenic mice expressing the human erythropoietin gene  

Energy Technology Data Exchange (ETDEWEB)

Erythropoietin is a glycoprotein hormone that regulates mammalian erythropoiesis. To study the expression of the human erythropoietin gene, EPO, 4 kilobases of DNA encompassing the gene with 0.4 kilobase of 5{prime} flanking sequence and 0.7 kilobase of 3{prime} flanking sequence was microinjected into fertilized mouse eggs. Transgenic mice were generated that are polycythemic, with increased erythrocytic indices in peripheral blood, increased numbers of erythroid precursors in hematopoietic tissue, and increased serum erythropoietin levels. Transgenic homozygotes show a greater degree of polycythemia than do heterozygotes as well as striking extramedullary erythropoiesis. Human erythropoietin RNA was found not only in fetal liver, adult liver, and kidney but also in all other transgenic tissues analyzed. Anemia induced increased human erythropoietin RNA levels in liver but not kidney. These transgenic mice represent a unique model of polycythemia due to increased erythropoietin levels.

Semenza, G.L.; Traystman, M.D.; Gearhart, J.D.; Antonarakis, S.E. (Johns Hopkins Univ. School of Medicine, Baltimore, MD (USA))

1989-04-01

304

Polycythemia in transgenic mice expressing the human erythropoietin gene  

International Nuclear Information System (INIS)

Erythropoietin is a glycoprotein hormone that regulates mammalian erythropoiesis. To study the expression of the human erythropoietin gene, EPO, 4 kilobases of DNA encompassing the gene with 0.4 kilobase of 5' flanking sequence and 0.7 kilobase of 3' flanking sequence was microinjected into fertilized mouse eggs. Transgenic mice were generated that are polycythemic, with increased erythrocytic indices in peripheral blood, increased numbers of erythroid precursors in hematopoietic tissue, and increased serum erythropoietin levels. Transgenic homozygotes show a greater degree of polycythemia than do heterozygotes as well as striking extramedullary erythropoiesis. Human erythropoietin RNA was found not only in fetal liver, adult liver, and kidney but also in all other transgenic tissues analyzed. Anemia induced increased human erythropoietin RNA levels in liver but not kidney. These transgenic mice represent a unique model of polycythemia due to increased erythropoietin levels

1989-01-01

305

Designer proton-channel transgenic algae for photobiological hydrogen production  

Energy Technology Data Exchange (ETDEWEB)

A designer proton-channel transgenic alga for photobiological hydrogen production that is specifically designed for production of molecular hydrogen (H.sub.2) through photosynthetic water splitting. The designer transgenic alga includes proton-conductive channels that are expressed to produce such uncoupler proteins in an amount sufficient to increase the algal H.sub.2 productivity. In one embodiment the designer proton-channel transgene is a nucleic acid construct (300) including a PCR forward primer (302), an externally inducible promoter (304), a transit targeting sequence (306), a designer proton-channel encoding sequence (308), a transcription and translation terminator (310), and a PCR reverse primer (312). In various embodiments, the designer proton-channel transgenic algae are used with a gas-separation system (500) and a gas-products-separation and utilization system (600) for photobiological H.sub.2 production.

Lee, James Weifu (Knoxville, TN)

2011-04-26

306

The Applications of Transgenic Rabbits in Agriculture and Biomedicine  

Directory of Open Access Journals (Sweden)

Full Text Available During the last decades transgenic rabbits have provided suitable biological model for regulating and manipulating of interest genes. Several studies showed transgenic rabbits can be produced by different methods. In the last two decades pronucleotide microinjection was conventional and common method for produce transgenic rabbits but the current studies demonstrated that SMGT method provides a simple and straightforward technology to introduce DNA into rabbits which has many advantages in comparison with other methods. The rabbit as both a laboratory and domestic animal species provides several opportunities for investigators to study the mechanisms of human disease such as lipoprotein metabolism, atherosclerosis and hypertrophic cardiomyopathy. Regarding to the present review transgenic rabbits in the last decade could have been managed to provide an alternative way to produce therapeutic proteins for treating human diseases.

Kh. Hosseini

2011-01-01

307

OPTIMAL BAND SELECTION OF HYPERSPECTRAL DATA FOR TRANSGENIC CORN IDENTIFICATION  

Science.gov (United States)

Resistance development by insect pests to the insecticidal proteins expressed in transgenic crops would increase reliance on broad spectrum chemical insecticides subsequently reducing environmental quality and increasing worker exposure to toxic chemicals. An important component ...

308

Transgenic Mouse Model For Development Of Cancer Therapeutics  

Science.gov (United States)

The National Cancer Institute's Medical Oncology Branch is seeking statements of capability or interest from parties interested in collaborative research to further develop, evaluate, or commercialize the thymidylate synthase transgenic mouse model.

309

Performance of Transgenic Bt Cotton Against Insect Pest Infestation  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Studies were carried out to investigate insect infestation on transgenic Bt and non-Bt cotton varieties and genotypes. The seeds of seven cotton varieties, viz., KMG-1, KMG-2, KMG-3, MS-1, MS-2 NIAB-78 and CRIS-134 were sown on May 15, 2002 in a completely randomized block design. The study comprised laboratory bioassay and field screening of different varieties and genotypes. Laboratory study indicated that transgenic Bt cotton was highly toxic to Earias vittella causing 100% larval m...

Abro, G. H.; Syed, T. S.; Tunio, G. M.; Khuhro, M. A.

2004-01-01

310

Optimised plasmids for sustained transgene expression in vivo  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Plasmid based gene therapy approaches often lack long term transgene expression in vivo due to silencing or loss of the vector. One way to overcome these limitations is to combine non-silenced promoters with strong viral enhancers. Here we combine cytomegalovirus (CMV) derived enhancer elements with the strong, human elongation factor 1 alpha (EF1a) promoter in a plasmid backbone devoid of potentially immunostimulating CpG sequences. The transgene expression of plasmids containing either the ...

2010-01-01

311

Analysis of the mouse protamine 1 promoter in transgenic mice.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Protamines are small arginine-rich proteins that package DNA in spermatozoa. The mouse protamine 1 (Prm-1) gene is transcribed exclusively in post-meiotic spermatids. To identify elements in the Prm-1 promoter required for spermatid-specific transcription, we generated transgenic mice by microinjection of transgenes containing Prm-1 5' flanking sequences with 5' truncations or internal deletions of conserved sequences linked to a marked Prm-1 gene. We also tested Prm-1 promoter regions with a...

Zambrowicz, B. P.; Harendza, C. J.; Zimmermann, J. W.; Brinster, R. L.; Palmiter, R. D.

1993-01-01

312

Generation and Characterization of Human Heme Oxygenase-1 Transgenic Pigs  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Xenotransplantation using transgenic pigs as an organ source is a promising strategy to overcome shortage of human organ for transplantation. Various genetic modifications have been tried to ameliorate xenograft rejection. In the present study we assessed effect of transgenic expression of human heme oxygenase-1 (hHO-1), an inducible protein capable of cytoprotection by scavenging reactive oxygen species and preventing apoptosis caused by cellular stress during inflammatory processes, in neon...

Yeom, Hye-jung; Koo, Ok Jae; Yang, Jaeseok; Cho, Bumrae; Hwang, Jong-ik; Park, Sol Ji; Hurh, Sunghoon; Kim, Hwajung; Lee, Eun Mi; Ro, Han; Kang, Jung Taek; Kim, Su Jin; Won, Jae-kyung; O Connell, Philip J.; Kim, Hyunil

2012-01-01

313

Transgenic Zebrafish Recapitulating tbx16 Gene Early Developmental Expression  

Digital Repository Infrastructure Vision for European Research (DRIVER)

We describe the creation of a transgenic zebrafish expressing GFP driven by a 7.5 kb promoter region of the tbx16 gene. This promoter segment is sufficient to recapitulate early embryonic expression of endogenous tbx16 in the presomitic mesoderm, the polster and, subsequently, in the hatching gland. Expression of GFP in the transgenic lines later in development diverges to some extent from endogenous tbx16 expression with the serendipitous result that one line expresses GFP specifically in co...

Wells, Simon; Nornes, Svanhild; Lardelli, Michael

2011-01-01

314

Efficient selection and evaluation of transgenic lines of Crambe abyssinica  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Crambe abyssinica is a dedicated oilseed crop suitable for production of industrial feedstocks. Genetic modification of crambe has progressed substantially in the last few years, but the transformation efficiency needs to be further improved. Meanwhile, developing a reliable molecular system including Southern blot and qRT-PCR analyses is desired for effectively evaluating transgenic lines and gene expression levels of both endogenous and transgenes. In this study, we have developed an effici...

Li, Xueyuan; Fan, Jing; Gruber, Jens; Guan, Rui; Frentzen, Margrit; Zhu, Li-hua

2013-01-01

315

Transgene x environment interactions in genetically modified wheat  

Digital Repository Infrastructure Vision for European Research (DRIVER)

BACKGROUND: The introduction of transgenes into plants may cause unintended phenotypic effects which could have an impact on the plant itself and the environment. Little is published in the scientific literature about the interrelation of environmental factors and possible unintended effects in genetically modified (GM) plants. METHODS AND FINDINGS: We studied transgenic bread wheat Triticum aestivum lines expressing the wheat Pm3b gene against the fungus powdery mildew Blumeria graminis f.sp...

Zeller, S. L.; Kalinina, O.; Brunner, S.; Keller, B.; Schmid, B.

2010-01-01

316

Spermatid-specific expression of protamine 1 in transgenic mice.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Protamines are abundant basic proteins involved in the condensation of sperm chromatin. In the mouse, protamine genes are transcribed postmeiotically in round spermatids. We have cloned and sequenced the mouse protamine 1 gene. Ten lines of transgenic mice harboring marked protamine 1 sequences were generated by microinjection of fertilized eggs. Transcription of the transgene is restricted to round spermatids and in several cases exceeds that of the endogenous gene. The cis-acting sequences ...

Peschon, J. J.; Behringer, R. R.; Brinster, R. L.; Palmiter, R. D.

1987-01-01

317

Pancreatic expression of human insulin gene in transgenic mice.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

We have investigated the possibility of obtaining integration and expression of a native human gene in transgenic mice. An 11-kilobase (kb) human chromosomal DNA fragment including the insulin gene (1430 base pairs) was microinjected into fertilized mouse eggs. This fragment was present in the genomic DNA of several developing animals. One transgenic mouse and its progeny were analyzed for expression of the foreign gene. Synthesis and release of human insulin was revealed by detection of the ...

Bucchini, D.; Ripoche, M. A.; Stinnakre, M. G.; Desbois, P.; Lore?s, P.; Monthioux, E.; Absil, J.; Lepesant, J. A.; Pictet, R.; Jami, J.

1986-01-01

318

Enhanced phytoremediation of volatile environmental pollutants with transgenic trees  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Small, volatile hydrocarbons, including trichloroethylene, vinyl chloride, carbon tetrachloride, benzene, and chloroform, are common environmental pollutants that pose serious health effects. We have developed transgenic poplar (Populus tremula × Populus alba) plants with greatly increased rates of metabolism and removal of these pollutants through the overexpression of cytochrome P450 2E1, a key enzyme in the metabolism of a variety of halogenated compounds. The transgenic poplar plants exh...

2007-01-01

319

Gone with transgenic cotton cropping in the USA  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The 2010 Beltwide Cotton Conferences provided a new vision of the consequences of about 15 years of widespread and uncoordinated cropping of transgenic cotton in the United States. Insect-resistant and/or herbicide-tolerant cotton varieties modified parasite complexes, namely those of insects and weeds damaging cotton crops. The Conferences have revealed that the adaptation solutions so far proposed make illusory the expectations at the launch of transgenic cotton, in terms of effective pest ...

Fok, Michel

2011-01-01

320

Enhancing shoot recovery from transgenic avocado somatic embryos  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Use of biotechnological tools in avocado (Persea americana Mill.) is hampered by difficulties in obtaining mature somatic embryos with an acceptable germination capacity. Use of semi-permeable cellulose acetate membranes on top of maturation medium has improved the quality of obtained embryos and their germination rate; however, in the case of transgenic embryos the conversion rate is still rather low. In this investigation, a protocol for recovery of transgenic plants has been developed. Mat...

Palomo-ri?os, Elena; Vidoy, Isabel; Barcelo?-mun?oz, Araceli; Mercado, Jose Angel; Pliego-alfaro, Fernando

2013-01-01

 
 
 
 
321

MicroRNA-Restricted Transgene Expression in the Retina  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Background: Gene transfer using adeno-associated viral (AAV) vectors has been successfully applied in the retina for the treatment of inherited retinal dystrophies. Recently, microRNAs have been exploited to fine-tune transgene expression improving therapeutic outcomes. Here we evaluated the ability of retinal-expressed microRNAs to restrict AAV-mediated transgene expression to specific retinal cell types that represent the main targets of common inherited blinding conditions. Methodology/Pri...

2011-01-01

322

Recombination technologies for enhanced transgene stability in bioengineered insects  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Transposon-based vectors currently provide the most suitable gene transfer systems for insect germ-line transformation and are used for molecular improvement of the Sterile Insect Technique. However, the long time stability of genome-integrated transposon constructs depends on the absence of transposase activity that could remobilize the transposon-embedded transgenes. To achieve transgene stability transposon vectors are usually non-autonomous, lacking a functional transposase gene, and chos...

2010-01-01

323

Use of Wolbachia to drive nuclear transgenes through insect populations.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Wolbachia is an inherited intracellular bacterium found in many insects of medical and economic importance. The ability of many strains to spread through populations using cytoplasmic incompatibility, involving sperm modification and rescue, provides a powerful mechanism for driving beneficial transgenes through insect populations, if such transgenes could be inserted into and expressed by Wolbachia. However, manipulating Wolbachia in this way has not yet been achieved. Here, we demonstrate t...

2004-01-01

324

Comprehensive Assessment of Milk Composition in Transgenic Cloned Cattle  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The development of transgenic cloned animals offers new opportunities for agriculture, biomedicine and environmental science. Expressing recombinant proteins in dairy animals to alter their milk composition is considered beneficial for human health. However, relatively little is known about the expression profile of the proteins in milk derived from transgenic cloned animals. In this study, we compared the proteome and nutrient composition of the colostrum and mature milk from three lines of ...

Zhang, Ran; Guo, Chengdong; Sui, Shunchao; Yu, Tian; Wang, Jianwu; Li, Ning

2012-01-01

325

Enhanced Malignant Tumorigenesis in Cdk4-Transgenic Mice  

Digital Repository Infrastructure Vision for European Research (DRIVER)

In a previous study, we reported that overexpression of CDK4 in mouse epidermis results in epidermal hyperplasia, hypertrophy and severe dermal fibrosis. In this study, we have investigated the susceptibility to skin tumor formation by forced expression of CDK4. Skin tumors from transgenic mice showed a dramatic increase in the rate of malignant progression to squamous cell carcinomas (SCC) in an initiation-promotion protocol. Histopathological analysis of papillomas from transgenic mice show...

Miliani Marval, Paula L.; Macias, Everardo; Conti, Claudio J.; Rodriguez-puebla, Marcelo L.

2004-01-01

326

A Primer for Using Transgenic Insecticidal Cotton in Developing Countries  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Many developing countries face the decision of whether to approve the testing and commercial use of insecticidal transgenic cotton and the task of developing adequate regulations for its use. In this review, we outline concepts and provide information to assist farmers, regulators and scientists in making decisions concerning this technology. We address seven critical topics: 1) molecular and breeding techniques used for the development of transgenic cotton cultivars, 2) properties of transge...

Showalter, Ann M.; Heuberger, Shannon; Tabashnik, Bruce E.; Carrie?re, Yves

2009-01-01

327

Local expression of transgene encoded TNF alpha in islets prevents autoimmune diabetes in nonobese diabetic (NOD) mice by preventing the development of auto-reactive islet-specific T cells  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Lately, TNF alpha has been the focus of studies of autoimmunity; its role in the progression of autoimmune diabetes is, however, still unclear. To analyze the effects of TNF alpha in insulin-dependent diabetes mellitus (IDDM), we have generated nonobese diabetic (NOD) transgenic mice expressing TNF alpha under the control of the rat insulin II promoter (RIP). In transgenic mice, TNF alpha expression on the islets resulted in massive insulitis, composed of CD4+ T cells, CD8+ T cells, and B cel...

1996-01-01

328

Genetic Quality Control of the Rat Strains at the National Bio Resource Project – Rat  

Directory of Open Access Journals (Sweden)

Full Text Available The National Bio Resource Project – Rat (NBRP-Rat comprises the largest bank of laboratory rat (Rattus norvegicus strains in the world. Its main focus is to develop infrastructure that will facilitate the systematic collection, preservation, and provision of rat strains. To breed effectively more than 180 rat strains in living stock, we establish the genetic control system in which a systematic set of genetic diagnoses and genetic monitoring are included. Genetic monitoring is performed by using 20 polymorphic markers. Monitoring is carried out when a living animal stock is re-established by using cryopreserved embryos or sperm or when a rat strain is first introduced to the NBRP-Rat by a depositor. Additional monitoring is then carried out on each strain every two years. Genetic diagnosis is performed largely by employing the Amp-FTA method. Protocols which detail how to perform a genetic diagnosis of 11 transgenes and 24 mutations have been made. Among the mutations, nine can be detected by simple gel electrophoresis of the PCR products, 11 by restriction enzyme treatment of the PCR products, and four by direct PCR product sequencing. Using this genetic control system, the NBRP-Rat can guarantee the genetic quality of its rat strains.

Tadao Serikawa

2010-11-01

329

Postlarval fitness of transgenic strains of Cochliomyia hominivorax (Diptera: Calliphoridae).  

Science.gov (United States)

Eight transgenic strains of Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae) were compared with the wild-type parental laboratory strain (P95) in colony. Measurements of average weight of pupae, percentage of adults emerging from pupae, ratio of males to total emerged adults, and mating competitiveness were analyzed. The parental strain colony was subcultured and exposed to handling procedures equivalent to transgenic strains for valid comparison of overall colony fitness. None of the transgenic colonies exhibited significantly lower fitness characteristics than the control parental colony. One transgenic colony had a higher ratio of adults emerging from pupae, and five colonies had higher average pupal weight; because fitness cost would only be indicated by lower values, the statistical variations were not significant. Males of one transgenic strain were shown to mate with equal frequency compared with males of the parental strain. Hence, the presence of the transgene used to produce the strains tested did not incur a fitness cost to the colonies of laboratory-reared C. hominivorax. PMID:15279308

Allen, Margaret L; Berkebile, Dennis R; Skoda, Steven R

2004-06-01

330

[Obtaining transgenic rice plants and their progenies using Agrobacterium tumefaciens].  

Science.gov (United States)

Rice (Oriza sativa L.) suspension cells of Taipei 309 were co-cultivated with A. tumefaciens stran EHA101 harbouring binary vector pBYT2 for 3 days in the presence of vir inducer, 100 mumol/L acetosyringone (AS). After 2 months of continuous selection, 17 stable hygromycin-resistant, GUS-positive calli were recovered from 364 suspension cell clusters co-cultivated with A. tumefaciens. 10 putative transgenic R0 plants obtained from 8 tansformed calli and their progenies were analyzed for the integration and expression of foreign genes. Southern blot analysis of R0 and R1 generations indicated that foreign genes had been stably integrated in the genome of transgenic rice and sexually transmitted. One of the transgenic lines showed 5 copies of T-DNA integration, while the others had only one copy. Histochemical staining observation and fluorometric assay of GUS activity in transgenic rice cells and plants showed ubiquitin promoter from maize was highly effective in driving the expression of gus reporter gene in transgenic rice cells. GUS protein and its activity were also investigated through ndPAGE-X-Gluc staining assay, and it was found that the GUS protein in transgenic rice cells was smaller in size than the standard GUS protein (Sigma Co. G0786) but as large as that from E.coli HB101 (pBI121). This study suggested that Agrobacterium-mediated transformation of plant is an efficient and reliable method to introduce foreign genes into rice. PMID:10465898

Yin, Z C; Yang, F; Xu, Y; Li, B J

1998-12-01

331

Non-Homologous End Joining Plays a Key Role in Transgene Concatemer Formation in Transgenic Zebrafish Embryos  

Digital Repository Infrastructure Vision for European Research (DRIVER)

This study focused on concatemer formation and integration pattern of transgenes in zebrafish embryos. A reporter plasmid based on enhanced green fluorescent protein (eGFP) driven by Cytomegalovirus (CMV) promoter, pCMV-pax6in-eGFP, was constructed to reflect transgene behavior in the host environment. After removal of the insertion fragment by double digestion with various combinations of restriction enzymes, linearized pCMV-pax6in-eGFP vectors were generated with different combinations of 5...

Dai, Jun; Cui, Xiaojuan; Zhu, Zuoyan; Hu, Wei

2010-01-01

332

Evaluating cerebellar functions using optogenetic transgenic mice  

Science.gov (United States)

We employed a transgenic mouse having conditional expression of ChR2(H134R) in neurons of the inferior olive to facilitate understanding of the role of electrical coupling and oscillation in central nervous system function. Two-photon excitation of ChR2-expressing neurons using 64 laser beams restricted to single inferior olive cell bodies depolarized neurons and evoked voltage deflections in neighboring neurons demonstrating electrical coupling. Broader illumination of neuronal ensembles using blue light induced an optical clamp of endogenous electrical rhythms in the inferior olive of acutely-prepared brain slices, which when applied in vivo directly modulated the local field potential activity and induced tremor. The experiments demonstrate novel methods to optically manipulate electrically coupled potentials and rhythmogenesis within a neuronal ensemble. From a functional perspective, the experiments shed light on the cellular and circuitry mechanisms of essential tremor, a prevalent neurological condition, by indicating time- and frequencydependence of tremor upon varying rhythms of inferior olive stimulation. The experiments indicate analog control of a brain rhythm that may be used to enhance our understanding of the functional consequences of central rhythmogenesis.

Welsh, John P.; Turecek, Josef; Turner, Eric E.

2013-03-01

333

Transgenic plants in the biopharmaceutical market.  

Science.gov (United States)

Many of our 'small-molecule-drugs' are natural products from plants, or are synthetic compounds based on molecules found naturally in plants. However, the vast majority of the protein therapeutics (or biopharmaceuticals) we use are from animal or human sources, and are produced commercially in microbial or mammalian bioreactor systems. Over the last few years, it has become clear that plants have great potential for the production of human proteins and other protein-based therapeutic entities. Plants offer the prospect of inexpensive biopharmaceutical production without sacrificing product quality or safety, and following the success of several plant-derived technical proteins, the first therapeutic products are now approaching the market. In this review, the different plant-based production systems are discussed and the merits of transgenic plants are evaluated compared with other platforms. A detailed discussion is provided of the development issues that remain to be addressed before plants become an acceptable mainstream production technology. The many different proteins that have already been produced using plants are described, and a sketch of the current market and the activities of the key players is provided. Despite the currently unclear regulatory framework and general industry inertia, the benefits of plant-derived pharmaceuticals are now bringing the prospect of inexpensive veterinary and human medicines closer than ever before. PMID:15757412

Twyman, Richard M; Schillberg, Stefan; Fischer, Rainer

2005-02-01

334

Enhanced polyhydroxybutyrate production in transgenic sugarcane.  

Science.gov (United States)

Polyhydroxybutyrate (PHB) is a bacterial polyester that has properties similar to some petrochemically produced plastics. Plant-based production has the potential to make this biorenewable plastic highly competitive with petrochemical-based plastics. We previously reported that transgenic sugarcane produced PHB at levels as high as 1.8% leaf dry weight without penalty to biomass accumulation, suggesting scope for improving PHB production in this species. In this study, we used different plant and viral promoters, in combination with multigene or single-gene constructs to increase PHB levels. Promoters tested included the maize and rice polyubiquitin promoters, the maize chlorophyll A/B-binding protein promoter and a Cavendish banana streak badnavirus promoter. At the seedling stage, the highest levels of polymer were produced in sugarcane plants when the Cavendish banana streak badnavirus promoter was used. However, in all cases, this promoter underwent silencing as the plants matured. The rice Ubi promoter enabled the production of PHB at levels similar to the maize Ubi promoter. The maize chlorophyll A/B-binding protein promoter enabled the production of PHB to levels as high as 4.8% of the leaf dry weight, which is approximately 2.5 times higher than previously reported levels in sugarcane. This is the first time that this promoter has been tested in sugarcane. The highest PHB-producing lines showed phenotypic differences to the wild-type parent, including reduced biomass and slight chlorosis. PMID:22369516

Petrasovits, Lars A; Zhao, Lihan; McQualter, Richard B; Snell, Kristi D; Somleva, Maria N; Patterson, Nii A; Nielsen, Lars K; Brumbley, Stevens M

2012-06-01

335

Enhanced neuroinflammation and pain hypersensitivity after peripheral nerve injury in rats expressing mutated superoxide dismutase 1  

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Full Text Available Abstract Background Neuroinflammation and nitroxidative stress are implicated in the pathophysiology of neuropathic pain. In view of both processes, microglial and astroglial activation in the spinal dorsal horn play a predominant role. The present study investigated the severity of neuropathic pain and the degree of glial activation in an inflammatory- and nitroxidative-prone animal model. Methods Transgenic rats expressing mutated superoxide dismutase 1 (hSOD1G93A are classically used as a model for amyotrophic lateral sclerosis (ALS. Because of the associated inflammatory- and nitroxidative-prone properties, this model was used to study thermal and mechanical hypersensitivity following partial sciatic nerve ligation (PSNL. Next to pain hypersensitivity assessment, microglial and astroglial activation states were moreover characterized, as well as inflammatory marker gene expression and the glutamate clearance system. Results PSNL induced thermal and mechanical hypersensitivity in both wild-type (WT and transgenic rats. However, the degree of thermal hypersensitivity was found to be exacerbated in transgenic rats while mechanical hypersensitivity was only slightly and not significantly increased. Microglial Iba1 expression was found to be increased in the ipsilateral dorsal horn of the lumbar spinal cord after PSNL but such Iba1 up-regulation was enhanced in transgenic rats as compared WT rats, both at 3 days and at 21 days after injury. Moreover, mRNA levels of Nox2, a key enzyme in microglial activation, but also of pro-inflammatory markers (IL-1? and TLR4 were not modified in WT ligated rats at 21 days after PSNL as compared to WT sham group while transgenic ligated rats showed up-regulated gene expression of these 3 targets. On the other hand, the PSNL-induced increase in GFAP immunoreactivity spreading that was evidenced in WT rats was unexpectedly found to be attenuated in transgenic ligated rats. Finally, GLT-1 gene expression and uptake activity were shown to be similar between WT sham and WT ligated rats at 21 days after injury, while both parameters were significantly increased in the ipsilateral dorsal region of the lumbar spinal cord of hSOD1G93A rats. Conclusions Taken together, our findings show that exacerbated microglial activation and subsequent inflammatory and nitroxidative processes are associated with the severity of neuropathic pain symptoms.

Lavand'homme Patricia

2011-04-01

336

Divergent Phenotypes in Mutant TDP-43 Transgenic Mice Highlight Potential Confounds in TDP-43 Transgenic Modeling  

Science.gov (United States)

The majority of cases of frontotemporal lobar degeneration and amyotrophic lateral sclerosis are pathologically defined by the cleavage, cytoplasmic redistribution and aggregation of TAR DNA binding protein of 43 kDa (TDP-43). To examine the contribution of these potentially toxic mechanisms in vivo, we generated transgenic mice expressing human TDP-43 containing the familial amyotrophic lateral sclerosis-linked M337V mutation and identified two lines that developed neurological phenotypes of differing severity and progression. The first developed a rapid cortical neurodegenerative phenotype in the early postnatal period, characterized by fragmentation of TDP-43 and loss of endogenous murine Tdp-43, but entirely lacking aggregates of ubiquitin or TDP-43. A second, low expressing line was aged to 25 months without a severe neurodegenerative phenotype, despite a 30% loss of mouse Tdp-43 and accumulation of lower molecular weight TDP-43 species. Furthermore, TDP-43 fragments generated during neurodegeneration were not C-terminal, but rather were derived from a central portion of human TDP-43. Thus we find that aggregation is not required for cell loss, loss of murine Tdp-43 is not necessarily sufficient in order to develop a severe neurodegenerative phenotype and lower molecular weight TDP-43 positive species in mouse models should not be inherently assumed to be representative of human disease. Our findings are significant for the interpretation of other transgenic studies of TDP-43 proteinopathy.

D'Alton, Simon; Altshuler, Marcelle; Cannon, Ashley; Dickson, Dennis W.; Petrucelli, Leonard; Lewis, Jada

2014-01-01

337

Hematological and biochemical indexes in blood of HBV transgenic mice  

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Full Text Available Objective To study the effects of gene integration of HBV on the biochemical and hematological indices in blood of transgenic mice.Methods The venous blood was collected from orbital venous plexus of 28 normal mice born in the same brood(HBsAg negative and 50 HBV transgenic mice(HBsAg positive,6-8 weeks in age.The blood routine examination was performed,including white blood cells(WBC,red blood cells(RBC,hemoglobin(Hb,platelets(PLT,lymphocyte percentage(L%,intermediate cell percentage(M%,the percentage of leaf cells(G% and blood biochemical parameters including glucose(GLU,urea nitrogen(BUN,creatinine(CREA,total protein(ALB,albumin(ALB,total bilirubin(TBIL,alanine aminotransferase(ALT,aspartate aminotransferase(AST,total cholesterol(CHOL and triglyceride(TG.The differential indexes were analyzed in HBsAg positive and negative mice,also the mice of different sex.Results Significant differences were found between the transgenic and normal mice in blood GLU,BUN,CREA,RBCs,Hb and PLT(P < 0.05,while no significant differences were found in other indices.No difference was found in the above 6 indices between the different sex of normal mice,while there was significant difference in blood GLU between males and females in transgenic mice,and it was higher in males than in females(P < 0.05.The blood GLU,CREA and BUN were higher in transgenic male mice than in normal male mice,and the contents of RBCs,Hb and PLT were higher in transgenic female mice than in normal female mice(P < 0.05.Conclusion HBV DNA may throw some influences on blood biochemical and hematological indexes of HBV transgenic mice,and it may provide some valuable references to the study of HBV pathogenesis in HBV animal models.

Feng-jiao ZHENG

2011-09-01

338

Hepatic Stellate Cell-Targeted Delivery of Hepatocyte Growth Factor Transgene via Bile Duct Infusion Enhances Its Expression at Fibrotic Foci to Regress Dimethylnitrosamine-Induced Liver Fibrosis  

Science.gov (United States)

Abstract Liver fibrosis generates fibrotic foci with abundant activated hepatic stellate cells and excessive collagen deposition juxtaposed with healthy regions. Targeted delivery of antifibrotic therapeutics to hepatic stellate cells (HSCs) might improve treatment outcomes and reduce adverse effects on healthy tissue. We delivered the hepatocyte growth factor (HGF) gene specifically to activated hepatic stellate cells in fibrotic liver using vitamin A–coupled liposomes by retrograde intrabiliary infusion to bypass capillarized hepatic sinusoids. The antifibrotic effects of DsRed2-HGF vector encapsulated within vitamin A–coupled liposomes were validated by decreases in fibrotic markers in vitro. Fibrotic cultures transfected with the targeted transgene showed a significant decrease in fibrotic markers such as transforming growth factor-?1. In rats, dimethylnitrosamine-induced liver fibrosis is manifested by an increase in collagen deposition and severe defenestration of sinusoidal endothelial cells. The HSC-targeted transgene, administered via retrograde intrabiliary infusion in fibrotic rats, successfully reduced liver fibrosis markers alpha-smooth muscle actin and collagen, accompanied by an increase in the expression of DsRed2-HGF near the fibrotic foci. Thus, targeted delivery of HGF gene to hepatic stellate cells increased the transgene expression at the fibrotic foci and strongly enhanced its antifibrotic effects.

Narmada, Balakrishnan Chakrapani; Kang, Yuzhan; Venkatraman, Lakshmi; Peng, Qiwen; Sakban, Rashidah Binte; Nugraha, Bramasta; Jiang, Xuan; Bunte, Ralph M.; So, Peter T.C.; Tucker-Kellogg, Lisa

2013-01-01

339

Digital gene expression analysis of mature seeds of transgenic maize overexpressing Aspergillus niger phyA2 and its non-transgenic counterpart.  

Science.gov (United States)

The next generation sequencing technologies have been recently used for transcriptome analysis in many organisms because of the decreased sequencing cost and increased sequence output. In this study, we used digital gene expression (DGE) technique to compare the transcriptomic changes in mature seeds between transgenic maize overexpressing Aspergillus niger phyA2 and its non-transgenic counterpart. Deep sequencing of DGE libraries of the transgenic and its non-transgenic counterpart seeds generated 3,783,500 and 3,790,500 reads of 21-nucleotide, respectively, with frequencies spanning over four orders of magnitude. In transgenic maize, 53.97% of the unambiguous signature tags were mapped to the maize B73 reference genome, and 46.47% of genes were detected by at least two reads; in non-transgenic maize, the corresponding numbers were 51.38% and 47.39%. Compared with non-transgenic counterpart, about 12% of detected genes were differentially expressed in the transcriptome of transgenic maize seeds. Among these differentially expressed genes, there were 23 transcription factors in 14 families and no allergen genes. Pathway enrichment analysis revealed that 21 pathways were significantly affected by the transgenic event, in which the pathway involved in protein processing in endoplasmic reticulum was the most significantly affected. Results from this study indicated that both intended and unintended transcriptomic changes occurred in the transgenic maize, thus emphasizing the importance of transcriptome profiling in risk assessment of transgenic events. PMID:23836108

Rao, Jun; Yang, Litao; Wang, Congmao; Zhang, Dabing; Shi, Jianxin

2013-01-01

340

Neutralizing antibodies against rotavirus produced in transgenically labelled purple tomatoes.  

Science.gov (United States)

Edible fruits are inexpensive biofactories for human health-promoting molecules that can be ingested as crude extracts or partially purified formulations. We show here the production of a model human antibody for passive protection against the enteric pathogen rotavirus in transgenically labelled tomato fruits. Transgenic tomato plants expressing a recombinant human immunoglobulin A (hIgA_2A1) selected against the VP8* peptide of rotavirus SA11 strain were obtained. The amount of hIgA_2A1 protein reached 3.6 ± 0.8% of the total soluble protein in the fruit of the transformed plants. Minimally processed fruit-derived products suitable for oral intake showed anti-VP8* binding activity and strongly inhibited virus infection in an in vitro virus neutralization assay. In order to make tomatoes expressing hIgA_2A1 easily distinguishable from wild-type tomatoes, lines expressing hIgA_2A1 transgenes were sexually crossed with a transgenic tomato line expressing the genes encoding Antirrhinum majus Rosea1 and Delila transcription factors, which confer purple colour to the fruit. Consequently, transgenically labelled purple tomato fruits expressing hIgA_2A1 have been developed. The resulting purple-coloured extracts from these fruits contain high levels of recombinant anti-rotavirus neutralizing human IgA in combination with increased amounts of health-promoting anthocyanins. PMID:22070155

Juárez, Paloma; Presa, Silvia; Espí, Joaquín; Pineda, Benito; Antón, María T; Moreno, Vicente; Buesa, Javier; Granell, Antonio; Orzaez, Diego

2012-04-01

 
 
 
 
341

Nutritional composition analysis of meat from human lactoferrin transgenic bulls.  

Science.gov (United States)

Transgenic technology has many potential advantages in food production. However, the transgenic technology process may influence the composition of food products derived from genetically engineered (GE) animals, which may be adverse to human health. Therefore, it is very important to research the compositions of GE animal products. Here, we analyzed the compositions of meat from the offspring of human lactoferrin (hLF) transgenic cows, which can express human lactoferrin proteins in their mammary gland. Six hLF transgenic bulls and three wide-type (WT) bulls, 10 months of age, were slaughtered for meat composition analysis. To determine the comparative health of hLF bulls for meat analysis, hematological analyses, organ/body weight analyses and pathology analyses were conducted. Results of the meat analysis show that there were no significant differences in the hematological parameters, organ/body weight ratios of hLF and WT bulls (P>0.05), and histopathological examination of the main organs of hLF bulls revealed no abnormalities. Nutrient parameters of meat compositions of hLF and WT bulls did not show any significant differences (P>0.05). All of these results suggest that the hLF transgene did not have an impact on the meat nutrient compositions of hLF bulls. PMID:23394369

Zhao, Jie; Xu, Jianxiang; Wang, Jianwu; Li, Ning

2013-01-01

342

Quality of transgenic laboratory strains of Cochliomyia hominivorax (Diptera: Calliphoridae).  

Science.gov (United States)

Genetically modified, mass reared insects present novel possibilities for the future of insect control. One concern about manipulation of insects is a possible loss of strain quality due to the introduction of a foreign gene of any sort into the insect genome. Eight transgenic strains of screwworm, Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae), were compared with the wild-type parental laboratory strain in laboratory culture. Measurements of average fertility, fecundity, larval productivity, and longevity were analyzed. Two transgenic strains had significantly lower larval productivity than controls, one of which was explained by a homozygous lethal insertion of the transgene. Another strain produced significantly fewer eggs than controls. Overall strain characteristics, including measurements from egg, larva, pupa, and adult stages, were compared. Transgenic colonies did not consistently show significantly lower individual or aggregate strain quality characteristics than the control parental colony; hence, the presence of the transgene used to produce the strains tested did not incur a discrete cost to the colonies of laboratory-reared C. hominivorax. PMID:16539163

Allen, Margaret L; Scholl, Philip J

2005-12-01

343

Handmade cloned transgenic piglets expressing the nematode fat-1 gene  

DEFF Research Database (Denmark)

Abstract Production of transgenic animals via somatic cell nuclear transfer (SCNT) has been adapted worldwide, but this application is somewhat limited by its relatively low efficiency. In this study, we used handmade cloning (HMC) established previously to produce transgenic pigs that express the functional nematode fat-1 gene. Codon-optimized mfat-1 was inserted into eukaryotic expression vectors, which were transferred into primary swine donor cells. Reverse transcriptase PCR (RT-PCR), gas chromatography, and chromosome analyses were performed to select donor clones capable of converting n-6 into n-3 fatty acids. Blastocysts derived from the clones that lowered the n-6/n-3 ratio to approximately 1:1 were transferred surgically into the uteri of recipients for transgenic piglets. By HMC, 37% (n=558) of reconstructed embryos developed to the blastocyst stage after 7 days of culture in vitro, with an average cell number of 81±36 (n=14). Three recipients became pregnant after 408 day-6 blastocysts were transferred into four naturally cycling females, and a total of 14 live offspring were produced. The nematode mfat-1 effectively lowered the n-6/n-3 ratio in muscle and major organs of the transgenic pig. Our results will help to establish a reliable procedure and an efficient option in the production of transgenic animals.

Zhang, Peng; Zhang, Yidi

2012-01-01

344

Reversible methylation and silencing of homologous transgenes in tobacco plants  

International Nuclear Information System (INIS)

Homology dependent gene silencing in transgenic plants occurs frequently when multiple copies of a transgene or a transgene with homology to an endogenous plant gene are present in a plant nucleus. The multiple copies can be arranged in cis on the same DNA molecule, or they can be present on different DNA molecules, either in allelic or non-allelic locations (trans inactivation). Although the phenomenon of silencing is well established, the mechanism by which it occurs are not completely understood. At present, different silencing systems can be distinguished by three major features: (1) the region of homology involved in the interaction (promoter or protein coding region); (2) the level at which silencing occurs (transcriptional or post-transcriptional); and (3) the degree of meiotic heritability of the silenced/methylated states after segregation of two interacting homologous loci in progeny. Interactions that lead to the silencing and methylation of partially homologous transgenes in tobacco have been studied. The transgenes share homology only in promoter regions; both the nopaline synthase promoter and the 35S promoter of the cauliflower mosaic virus have been used to drive the expression of various selectable and biochemical marker genes. The properties of these silencing systems are discussed, with particular reference to the features mentioned above. (author). 13 refs

1995-11-01

345

Identification of abnormal gene expression in bovine transgenic somatic cell nuclear transfer embryos.  

Science.gov (United States)

This study was conducted to investigate the expression of three genes related to early embryonic development in bovine transgenic cloned embryos. To accomplish this, development of bovine transgenic somatic cell nuclear transfer (SCNT) embryos was compared with non-transgenic embryos. Next, mRNA transcription of three specific genes (DNMT1, Hsp 70.1, and Mash2) related to early embryo development in transgenic SCNT embryos was compared between transgenic and non-transgenic SCNTs, parthenogenetic embryos, and in vitro fertilization (IVF) embryos. Transgenic SCNT embryos showed significantly lower rates of development to the blastocyst stage than non-transgenic ones. To investigate normal gene expression, RNA was extracted from ten blastocysts derived from parthenogenesis, IVF, non-transgenic, and transgenic SCNT embryos and reverse-transcribed to synthesize cDNA. The cDNA was then subjected to PCR amplification and semi-quantified. More DNMT1 mRNA was detected in the transgenic SCNT group than the other three groups. Hsp 70.1 mRNA was detected in the IVF embryos, while lower levels were found in SCNT and parthenogenetic embryos. Mash2 mRNA was present at the highest levels in transgenic SCNT embryos. In conclusion, the higher levels of methylation and lower protein synthesis after heat shock in the transgenic SCNT embryos expected based on our results may cause lower embryonic development. PMID:24675837

Cho, Jongki; Kang, Sungkeun; Lee, Byeong Chun

2014-06-01

346

Faster Recovery of Cerebral Perfusion in SOD1-Overexpressed Rats after Cardiac Arrest and Resuscitation  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Protracted hypoperfusion is one of the hallmarks of secondary cerebral derangement after cardiac arrest and resuscitation (CAR), and reactive oxygen species have been implicated in reperfusion abnormalities. Using transgenic (Tg) rats overexpressing copper zinc superoxide dismutase (SOD1), we investigated the role of this intrinsic antioxidant in the restoration of cerebral blood flow (CBF) after CAR. Nine Tg and 11 wild-type (WT) rats were subjected to 14-min cardiac arrest, and CBF was meas...

Xu, Y.; Liachenko, S. M.; Tang, P.; Chan, P. H.

2009-01-01

347

[Nuclear transfer of goat somatic cells transgenic for human lactoferrin].  

Science.gov (United States)

Transgenic animal mammary gland bioreactors are being used to produce recombinant proteins with appropriate post-translational modifications, and nuclear transfer of transgenic somatic cells is a more powerful method to produce mammary gland bioreactor. Here we describe efficient gene transfer and nuclear transfer in goat somatic cells. Gene targeting vector pGBC2LF was constructed by cloning human lactoferrin (LF) gene cDNA into exon 2 of the milk goat beta-casein gene, and the endogenous start condon was replaced by that of human LF gene. Goat fetal fibroblasts were transfected with linearized pGBC2LF and 14 cell lines were positive according to PCR and Southern blot. The transgenic cells were used as donor cells of nuclear transfer, and some of reconstructed embryos could develop to blastocyst in vitro. PMID:17138536

Li, Lan; Shen, Wei; Pan, Qing-Yu; Min, Ling-Jiang; Sun, Yu-Jiang; Fang, Yong-Wei; Deng, Ji-Xian; Pan, Qing-Jie

2006-12-01

348

Bioassay for detection of transgenic soybean seeds tolerant to glyphosate  

Directory of Open Access Journals (Sweden)

Full Text Available Glyphosate is a systemic, nonselective, postemergence herbicide that inhibits growth of both weeds and crop plants. Once inside the plant, glyphosate interferes with biosynthesis of aromatic amino acids phenylalanine, tyrosine, and tryptophan, by inhibiting the activity of 5enolpyruvylshikimate-3-phosphate synthase (EPSPS, a key enzyme of the shikimate pathway. The objective of this work was to develop a simple, effective and inexpensible method for identification of transgenic soybean tolerant to glyphosate. This technique consisted in germinating soybean seeds in filter paper moistened with 100 to 200 muM of glyphosate. Transgenic soybean seeds tolerant to glyphosate germinated normally in this solution and, between 7 and 10 days, started to develop a primary root system. However non-transgenic seeds stopped primary root growth and emission of secondary roots.

Torres Antonio Carlos

2003-01-01

349

Identification of transgenic foods using NIR spectroscopy: A review  

Science.gov (United States)

The utilization of chemometric methods in the quantitative and qualitative analysis of feeds, foods, medicine and so on has been accompanied with the great evolution in the progress and in the near infrared spectroscopy (NIRS). Hence, recently the application of NIR spectroscopy has extended on the context of genetics and transgenic products. The aim of this review was to investigate the application of NIR spectroscopy to identificate transgenic products and to compare it with the traditional methods. The results of copious researches showed that the application of NIRS technology was successful to distinguish transgenic foods and it has advantages such as fast, avoiding time-consuming, non-destructive and low cost in relation to the antecedent methods such as PCR and ELISA.

Alishahi, A.; Farahmand, H.; Prieto, N.; Cozzolino, D.

2010-01-01

350

Establishment and detection of HBV transgenic mice with YMDD mutation  

Directory of Open Access Journals (Sweden)

Full Text Available Objective To establish the hepatitis B virus(HBV transgenic mice with YMDD mutation,and provide an animal model for research of HBV prevention and therapeutic approach.Methods 1.3 copies HBV genome containing YMDD mutation associated with lamivudine resistance was injected into the zygote of FVB/N female mice by microinjection.Integration and passage of exogenous gene in transgenic mice was confirmed by PCR.The expression of HBsAg in liver and kidney tissues in transgenic mice was identified by ELISA and immunohistochemistry.Results A total of 3401 zygotes were injected and 269 F0 pups were born.PCR analysis indicated that 33 out of 269 pups were positive,and the integrating rate of exogenous gene was 12.3% in F0.Fluorescent quantitative PCR showed that HBV DNA was weakly positive in serum samples in 9 transgenic mice,less than 103copies/ml.The expression of HBsAg in transgenic mice was observed in liver and kidney tissues by immunohistochemistry,and it was higher in kidney than in liver.The target gene was detected by PCR in 27.6% of 47 F1 offsprings.The expression of HBsAg could be observed in liver and kidney tissues in F1,which was similar to that in F0.Conclusion 1.3 copies HBV transgenic mice model containing YMDD mutation associated with lamivudine resistance is successfully produced by microinjection,and HBsAg expression can be transmitted through germline cells.

Yu-qin YOU

2011-09-01

351

A set of highly informative rat simple sequence length polymorphism (SSLP markers and genetically defined rat strains  

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Full Text Available Abstract Background The National Bio Resource Project for the Rat in Japan (NBRP-Rat is focusing on collecting, preserving and distributing various rat strains, including spontaneous mutant, transgenic, congenic, and recombinant inbred (RI strains. To evaluate their value as models of human diseases, we are characterizing them using 109 phenotypic parameters, such as clinical measurements, internal anatomy, metabolic parameters, and behavioral tests, as part of the Rat Phenome Project. Here, we report on a set of 357 simple sequence length polymorphism (SSLP markers and 122 rat strains, which were genotyped by the marker set. Results The SSLP markers were selected according to their distribution patterns throughout the whole rat genome with an average spacing of 7.59 Mb. The average number of informative markers between all possible pairs of strains was 259 (72.5% of 357 markers, showing their high degree of polymorphism. From the genetic profile of these rat inbred strains, we constructed a rat family tree to clarify their genetic background. Conclusion These highly informative SSLP markers as well as genetically and phenotypically defined rat strains are useful for designing experiments for quantitative trait loci (QTL analysis and to choose strategies for developing new genetic resources. The data and resources are freely available at the NBRP-Rat web site 1.

Yamasaki Ken-ichi

2006-04-01

352

A new highly effective anticysticercosis vaccine expressed in transgenic papaya.  

Science.gov (United States)

The use of transgenic plants as new antigen-delivery systems for subunit vaccines has been increasingly explored. We herein report progress toward a papaya-based vaccine against cysticercosis. Synthetic peptides (KETc1, KETc12, KETc7) were successfully expressed in 19 different transgenic papaya clones and found to be immunogenic. Complete protection against cysticercosis was induced with the soluble extract of the clones that expressed the higher levels of transcripts in up to 90% of the immunized mice. This study represents a key step towards the development of a more effective, sustainable and affordable oral subunit vaccine against human and pig cysticercosis. PMID:17399859

Hernández, Marisela; Cabrera-Ponce, José Luis; Fragoso, Gladis; López-Casillas, Fernando; Guevara-García, Arturo; Rosas, Gabriela; León-Ramírez, Claudia; Juárez, Patricia; Sánchez-García, Guadalupe; Cervantes, Jaquelynne; Acero, Gonzalo; Toledo, Andrea; Cruz, Carmen; Bojalil, Rafael; Herrera-Estrella, Luis; Sciutto, Edda

2007-05-22

353

Developmental analysis of the cytomegalovirus enhancer in transgenic animals.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The major immediate-early promoter (MIEP) of human, cytomegalovirus (HCMV) constitutes a primary genetic switch for viral activation. In this study, regulation of the enhancer-containing segment (nucleotides -670 to +54) of the HCMV MIEP attached to the 1acZ reporter gene was examined in the developing embryos of transgenic mice to identify temporal and tissue-specific expression. We find that the transgene reporter is first detected as a dorsal stripe of expression in the neural folds of emb...

Baskar, J. F.; Smith, P. P.; Ciment, G. S.; Hoffmann, S.; Tucker, C.; Tenney, D. J.; Colberg-poley, A. M.; Nelson, J. A.; Ghazal, P.

1996-01-01

354

Transient development of ovotestes in XX Sox9 transgenic mice  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The sex of an individual results from the paternal transmission of the SRY gene located on the Y chromosome. In turn, SRY initiates Sox9 expression, a transcription factor required for testicular differentiation. Ectopic activation of SOX9 in XX Wt1:Sox9 transgenic mice, induces female-to-male sex reversal in adult mice. Here we show that complete sex reversal is preceded by a transient phase of ovotestis differentiation with XX Wt1:Sox9 transgenic gonads containing a testicular central regio...

Gregoire, Elodie P.; Lavery, Rowena; Chassot, Anne-amandine; Akiyama, Haruhiko; Treier, Mathias; Behringer, Richard R.; Chaboissier, Marie-christine

2011-01-01

355

Biodiversity versus transgenic sugar beet: the one euro question  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The decision on whether to release transgenic crops in the EU is subject to irreversibility, uncertainty and flexibility. We analyse the case of herbicide-tolerant sugar beet and assess whether the EU’s 1998 de facto moratorium on transgenic crops for sugar beet was correct from a cost–benefit perspective, using a real option approach. We show that the decision was correct, providing households on average value the possible annual irreversible costs of herbicide-tolerant sugar beet at 1 o...

Demont, M.; Wesseler, J. H. H.; Tollens, E.

2004-01-01

356

Transgenic mouse model for the fragile X syndrome  

Energy Technology Data Exchange (ETDEWEB)

Transgenic fragile X knockout mice have been constructed to provide an animal model to study the physiologic function of the fragile X gene (FMR1) and to gain more insight into the clinical phenotype caused by the absence of the fragile X protein. Initial experiments suggested that the knockout mice show macroorchidism and cognitive and behavioral deficits, abnormalities comparable to those of human fragile X patients. In the present study, we have extended our experiments, and conclude that the Fmr1 knockout mouse is a reliable transgenic model to study the fragile X syndrome. 20 refs., 2 figs., 1 tab.

Kooy, R.F.; Reyniers, E.; De Boulle, K. [Univ. of Antwerp (Belgium)] [and others

1996-08-09

357

Transgenic mouse model for the fragile X syndrome.  

Science.gov (United States)

Transgenic fragile X knockout mice have been constructed to provide an animal model to study the physiologic function of the fragile X gene (FMR1) and to gain more insight into the clinical phenotype caused by the absence of the fragile X protein. Initial experiments suggested that the knockout mice show macroorchidism and cognitive and behavioral deficits, abnormalities comparable to those of human fragile X patients. In the present study, we have extended our experiments, and conclude that the Fmr1 knockout mouse is a reliable transgenic model to study the fragile X syndrome. PMID:8844056

Kooy, R F; D'Hooge, R; Reyniers, E; Bakker, C E; Nagels, G; De Boulle, K; Storm, K; Clincke, G; De Deyn, P P; Oostra, B A; Willems, P J

1996-08-01

358

The maintenance of methylation-free islands in transgenic mice.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The Thy-1 gene is expressed in a tissue- and stage-specific pattern and has a typical 1.6kb methylation-free island (MFI) covering about 600bp upstream and downstream of the two alternative first exons. By microinjection of a mouse Thy-1.1/human Thy-1 gene into fertilized eggs, we were able to show that the MFI is restored in the transgenic mice. The flanking sequence became methylated, but the MFI remains unmethylated in all tissues of transgenic mice at different developmental stages tested...

Kolsto, A. B.; Kollias, G.; Giguere, V.; Isobe, K. I.; Prydz, H.; Grosveld, F.

1986-01-01

359

Differential subcellular targeting of recombinant human ??-proteinase inhibitor influences yield, biological activity and in planta stability of the protein in transgenic tomato plants.  

Science.gov (United States)

The response of protein accumulation site on yield, biological activity and in planta stability of therapeutic recombinant human proteinase inhibitor (??-PI) was analyzed via targeting to different subcellular locations, like endoplasmic reticulum (ER), apoplast, vacuole and cytosol in leaves of transgenic tomato plants. In situ localization of the recombinant ??-PI protein in transgenic plant cells was monitored by immunohistochemical staining. Maximum accumulation of recombinant ??-PI in T? and T? transgenic tomato plants was achieved from 1.5 to 3.2% of total soluble protein (TSP) by retention in ER lumen, followed by vacuole and apoplast, whereas cytosolic targeting resulted into degradation of the protein. The plant-derived recombinant ??-PI showed biological activity for elastase inhibition, as monitored by residual porcine pancreatic elastase (PPE) activity assay and band-shift assay. Recombinant ??-PI was purified from transgenic tomato plants with high yield, homogeneity and biological activity. Purified protein appeared as a single band of ?48-50 kDa on SDS-PAGE with pI value ranging between 5.1 and 5.3. Results of mass spectrometry and optical spectroscopy of purified recombinant ??-PI revealed the structural integrity of the recombinant protein comparable to native serum ??-PI. Enzymatic deglycosylation and lectin-binding assays with the purified recombinant ??-PI showed compartment-specific N-glycosylation of the protein targeted to ER, apoplast and vacuole. Conformational studies based on urea-induced denaturation and circular dichroism (CD) spectroscopy revealed relatively lower stability of the recombinant ??-PI protein, compared to its serum counterpart. Pharmacokinetic evaluation of plant derived recombinant and human plasma-purified ??-PI in rat, by intravenous route, revealed significantly faster plasma clearance and lower area under curve (AUC) of recombinant protein. Our data suggested significance of protein sorting sequences and feasibility to use transgenic plants for the production of stable, glycosylated and biologically active recombinant ??-PI for further therapeutic applications. PMID:23017899

Jha, Shweta; Agarwal, Saurabh; Sanyal, Indraneel; Jain, G K; Amla, D V

2012-11-01

360

Overexpression of Glyoxalase-I Reduces Hyperglycemia-induced Levels of Advanced Glycation End Products and Oxidative Stress in Diabetic Rats*  

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The reactive advanced glycation end product (AGE) precursor methylglyoxal (MGO) and MGO-derived AGEs are associated with diabetic vascular complications and also with an increase in oxidative stress. Glyoxalase-I (GLO-I) transgenic rats were used to explore whether overexpression of this MGO detoxifying enzyme reduces levels of AGEs and oxidative stress in a rat model of diabetes. Rats were made diabetic with streptozotocin, and after 12 weeks, plasma and multiple tissues were isolated for an...

Brouwers, Olaf; Niessen, Petra M.; Ferreira, Isabel; Miyata, Toshio; Scheffer, Peter G.; Teerlink, Tom; Schrauwen, Patrick; Brownlee, Michael; Stehouwer, Coen D.; Schalkwijk, Casper G.

2011-01-01

 
 
 
 
361

Detection of transgenic cp4 epsps genes in the soil food web  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The persistence and movement of transgenic DNA in agricultural and natural systems is largely unknown. This movement poses a threat of horizontal gene transfer and possible proliferation of genetically modified DNA into the general environment. To assess the persistence of transgenic DNA in a field of Roundup Ready® corn, we quantified the presence of the transgene for glyphosate tolerance within a soil food web. Using quantitative real-time PCR, we identified the cp4 epsps transgene in bulk...

Hart, Miranda M.; Powell, Jeff R.; Gulden, Robert H.; Levy-booth, David J.; Dunfield, Kari E.; Peter Pauls, K.; Swanton, Clarence J.; Klironomos, John N.; Trevors, Jack T.

2009-01-01

362

Neuron-Specific Activation of Murine Cytomegalovirus Early Gene e1 Promoter in Transgenic Mice  

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The brain is the main target in congenital cytomegalovirus (CMV) infection and immunocompromised patients. No definite evidence that a CMV has special affinity for the central nervous system (CNS) has been published. Here, we generated transgenic mice with an e1 promoter/enhancer region connected to the reporter gene lacZ. Surprisingly, expression of the transgene was completely restricted to the CNS in all lines of transgenic mice. The transgene was expressed in subpopulation of neurons in t...

2003-01-01

363

REGULATED EXPRESSION OF TRANSGENES IN EMBRYONIC STEM CELL-DERIVED NEURAL CELLS  

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Discovery and characterization of gene promoters, enhancers and repressor binding elements is an important research area in neuroscience. Here, the suitability of embryonic stem cells and their neural derivatives as a model system for this research is investigated. Three neural transgenic constructs (from the Mnx1, Fabp7, and tuba1a genes) that have been validated in transgenic mice were inserted into embryonic stem cells as stable transgenes. These transgenic embryonic stem cells were differ...

2011-01-01

364

Allelic exclusion in transgenic mice carrying mutant human IgM genes  

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Expression of the membrane-bound version of the human mu chain in transgenic mice results in the allelic exclusion of endogenous mouse Ig heavy chain genes (6). The secreted version of the human Ig transgene has no such effect. F1 hybrid animals that carry transgenes for both secreted and membrane-bound human mu chains produce both forms of the human heavy chain while strongly suppressing endogenous mouse mu expression. The simultaneous expression of the two rearranged transgenes in primary B...

1988-01-01

365

A transgenic mouse that expresses a diversity of human sequence heavy and light chain immunoglobulins.  

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We have generated transgenic mice that express a diverse repertoire of human sequence immunoglobulins. The expression of this repertoire is directed by light and heavy chain minilocus transgenes comprised of human protein coding sequences in an unrearranged, germ-line configuration. In this paper we describe the construction of these miniloci and the composition of the CDR3 repertoire generated by the transgenic mice. The largest transgene discussed is a heavy chain minilocus that includes hu...

Taylor, L. D.; Carmack, C. E.; Schramm, S. R.; Mashayekh, R.; Higgins, K. M.; Kuo, C. C.; Woodhouse, C.; Kay, R. M.; Lonberg, N.

1992-01-01

366

Tolerance induced by physiological levels of secreted proteins in transgenic mice expressing human insulin.  

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We have used transgenic mice to study immune tolerance to autologous, non-MHC encoded proteins that are expressed at physiological levels in the circulation. The transgenic mice used in these studies express the human preproinsulin gene and synthesize human proinsulin. Human and mouse insulin are secreted from the pancreatic islets of transgenic mice in response to normal physiological stimuli, such as glucose. Our data demonstrate that the transgenic mice have acquired tolerance to human ins...

Whiteley, P. J.; Lake, J. P.; Selden, R. F.; Kapp, J. A.

1989-01-01

367

Evaluating the fitness of human lysozyme transgenic dairy goats: growth and reproductive traits  

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While there are many reports in the literature describing the attributes of specific applications of transgenic animals for agriculture, there are relatively few studies focusing on the fitness of the transgenic animals themselves. This work was designed to gather information on genetically modified food animals to determine if the presence of a transgene can impact general animal production traits. More specifically, we used a line of transgenic dairy goats expressing human lysozyme in their...

Jackson, Kathryn A.; Berg, Jolene M.; Murray, James D.; Maga, Elizabeth A.

2010-01-01

368

Overexpression of apolipoprotein AII in transgenic mice converts high density lipoproteins to proinflammatory particles.  

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Previous studies showed that transgenic mice overexpressing either apolipoprotein AI (apoAI) or apolipoprotein AII (apoAII), the major proteins of HDL, exhibited elevated levels of HDL cholesterol, but, whereas the apoAI-transgenic mice were protected against atherosclerosis, the apoAII-transgenic mice had increased lesion development. We now examine the basis for this striking functional heterogeneity. HDL from apoAI transgenics exhibited an enhanced ability to promote cholesterol efflux fro...

Castellani, L. W.; Navab, M.; Lenten, B. J.; Hedrick, C. C.; Hama, S. Y.; Goto, A. M.; Fogelman, A. M.; Lusis, A. J.

1997-01-01

369

Introducing Transgenes Into Insect Populations Using Combined Gene-drive Strategies: Modeling and Analysis  

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Engineered underdominance (EU), meiotic drive (MD) and Wolbachia have been proposed as mechanisms for driving anti-pathogen transgenes into natural populations of insect vectors of human diseases. EU can drive transgenes to high and stable frequencies but requires the release of sizeable numbers of engineered insects. MD and Wolbachia either cannot maintain high frequencies of transgenes or lack appropriate expression in critical tissues, but both can drive the transgenes to spread from very ...

2007-01-01

370

Male mating strategy and the introgression of a growth hormone transgene  

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Escaped transgenic organisms (GMO's) may threaten the populations of their wild relatives if able to hybridize with each other. The introgression of a growth enhancement transgene into a wild Atlantic salmon population may be affected by the transgene's effects not only on fitness parameters, but also on mating behaviour. Large anadromous GMO males are most preferred in mating, but a transgene can also give the large sneakers a reproductive advantage over the smaller wild individuals. With a ...

2008-01-01

371

Inducible expression of RANKL in transgenic pigs under the control of the Tet-On system  

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Because of the tremendous need for transgenic large animal models for human diseases, the process of SCNT is a crucial step in transgenic pig production. In our study, we evaluated the particular steps during the production for their impact on the efficiency of cloning transgenic pigs. For this purpose, statistical analysis was performed for all SCNT data from the years 2006 until June 2010. The RANKL transgenic osteoporosis model was chosen for an example for the production steps needed to f...

Schilling, Eleonore

2011-01-01

372

Growth hormone transgenic salmon pay for growth potential with increased predation mortality.  

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Recent advances in gene technology have been applied to create fast-growing transgenic fish, which are of great commercial interest owing to their potential to shorten production cycles and increase food production. However, there is growing concern and speculation over the impact that escaped growth hormone (GH)-transgenic fish may have on the natural environment. To predict these risks it is crucial to obtain empirical data on the relative fitness of transgenic and non-transgenic fish under...

2004-01-01

373

Dominos in the dairy: An analysis of transgenic maize in Dutch dairy farming  

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EU member states require farmers growing transgenic maize to respect a minimum distance from fields with non-transgenic maize. Previous studies have theoretically argued that such minimum distance requirements may lead to a so-called ‘domino effect’ where farmers who want to grow transgenic maize are forced to grow the non-transgenic variety and in turn impose the same constraints on their neighbors. This article applies a spatially explicit farm model to a dairy region in the Southern Ne...

Groeneveld, R. A.; Wesseler, J. H. H.; Berentsen, P. B. M.

2013-01-01

374

Rat skeleton  

Science.gov (United States)

Rats have low profile bodies that allow them to fit into small spaces to hide from predators. Rats use their sharp teeth and claws to break through bags, boxes, etc. to find food. Rats will eat whatever food is available, which means they are scavengers.

Katie Hale (CSUF;)

2007-09-01

375

Transgenic mice expressing green fluorescent protein under the control of the melanocortin-4 receptor promoter.  

Science.gov (United States)

The melanocortin-4 receptor (MC4-R) is an important regulator of energy homeostasis, and evidence suggests that MC4-R-expressing neurons are downstream targets of leptin action. MC4-Rs are broadly expressed in the CNS, and the distribution of MC4-R mRNA has been analyzed most extensively in the rat. However, relatively little is known concerning chemical profiles of MC4-R-expressing neurons. The extent to which central melanocortins act presynaptically or postsynaptically on MC4-Rs is also unknown. To address these issues, we have generated a transgenic mouse line expressing green fluorescent protein (GFP) under the control of the MC4-R promoter, using a modified bacterial artificial chromosome. We have confirmed that the CNS distribution of GFP-producing cells is identical to that of MC4-R mRNA in wild-type mice and that nearly all GFP-producing cells coexpress MC4-R mRNA. For example, cells coexpressing GFP and MC4-R mRNA were distributed in the paraventricular hypothalamic nucleus (PVH) and the dorsal motor nucleus of the vagus (DMV). MC4-R promotor-driven GFP expression was found in PVH cells producing thyrotropin-releasing hormone and in cholinergic DMV cells. Finally, we have observed that a synthetic MC3/4-R agonist, MT-II, depolarizes some GFP-expressing cells, suggesting that MC4-Rs function postsynaptically in some instances and may function presynaptically in others. These studies extend our knowledge of the distribution and function of the MC4-R. The transgenic mouse line should be useful for future studies on the role of melanocortin signaling in regulating feeding behavior and autonomic homeostasis. PMID:12904474

Liu, Hongyan; Kishi, Toshiro; Roseberry, Aaron G; Cai, Xiaoli; Lee, Charlotte E; Montez, Jason M; Friedman, Jeffrey M; Elmquist, Joel K

2003-08-01

376

Utilization of transgenic models in the evaluation of osteogenic differentiation of embryonic stem cells.  

Science.gov (United States)

Previous studies reported that embryonic stem cells (ESCs) can be induced to differentiate into cells showing a mature osteoblastic phenotype by culturing them under osteo-inductive conditions. It is probable that osteogenic differentiation requires that ESCs undergo differentiation through an intermediary step involving a mesenchymal lineage precursor. Based on our previous studies indicating that adult mesenchymal progenitor cells express ?-smooth muscle actin (?SMA), we have generated ESCs from transgenic mice in which an ?SMA promoter directs the expression of red fluorescent protein (RFP) to mesenchymal progenitor cells. To track the transition of ESC-derived MSCs into mature osteoblasts, we have utilized a bone-specific fragment of rat type I collagen promoter driving green fluorescent protein (Col2.3GFP). Following osteogenic induction in ESCs, we have observed expression of alkaline phosphatase (ALP) and subsequent mineralization as detected by von Kossa staining. After 1 week of osteogenic induction, ESCs begin to express ?SMARFP. This expression was localized to the peripheral area encircling a typical ESC colony. Nevertheless, these ?SMARFP positive cells did not show activation of the Col2.3GFP promoter, even after 7 weeks of osteogenic differentiation in vitro. In contrast, Col2.3GFP expression was detected in vivo, in mineralized areas following teratoma formation. Our results indicate that detection of ALP activity and mineralization of ESCs cultured under osteogenic conditions is not sufficient to demonstrate osteogenic maturation. Our study indicates the utility of the promoter-visual transgene approach to assess the commitment and differentiation of ESCs into the osteoblast lineage. PMID:23782451

Repic, Dario; Torreggiani, Elena; Franceschetti, Tiziana; Matthews, Brya G; Ivcevic, Sanja; Lichtler, Alexander C; Grcevic, Danka; Kalajzic, Ivo

2013-01-01

377

Utilization of transgenic models in evaluation of osteogenic differentiation of embryonic stem cells  

Science.gov (United States)

Previous studies reported that embryonic stem cells (ESCs) can be induced to differentiate into cells showing a mature osteoblastic phenotype by culturing them under osteo-inductive conditions. It is probable that osteogenic differentiation requires that ESCs undergo differentiation through an intermediary step involving a mesenchymal lineage precursor. Based on our previous studies indicating that adult mesenchymal progenitor cells express ?SMA, we have generated ESCs from transgenic mice in which an ?SMA promoter directs the expression of red fluorescent protein (RFP) to mesenchymal progenitor cells. To track the transition of ESC-derived MSCs into mature osteoblasts, we have utilized a bone-specific fragment of rat type I collagen promoter driving green fluorescent protein (Col2.3GFP). Following osteogenic induction in ESCs, we have observed expression of alkaline phosphatase and subsequent mineralization as detected by von Kossa staining. After one week of osteogenic induction, ESCs begin to express ?SMARFP. This expression was localized to the peripheral area encircling a typical ESC colony. Nevertheless, these ?SMARFP positive cells did not show activation of the Col2.3GFP promoter, even after 7 weeks of osteogenic differentiation in vitro. In contrast, Col2.3GFP expression was detected in vivo, in mineralized areas following teratoma formation. Our results indicate that detection of alkaline phosphatase activity and mineralization of ESCs cultured under osteogenic conditions is not sufficient to demonstrate osteogenic maturation. Our study indicates the utility of the promoter-visual transgene approach to assess the commitment and differentiation of ESCs into the osteoblast lineage.

Repic, Dario; Torreggiani, Elena; Franceschetti, Tiziana; Matthews, Brya G.; Ivcevic, Sanja; Lichtler, Alexander C.; Grcevic, Danka; Kalajzic, Ivo

2014-01-01

378

Maintenance and distribution of transgenic mice susceptible to human viruses: memorandum from a WHO meeting.  

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This Memorandum discusses the use of transgenic mice in poliovirus research and the potential risks to public health. General and specific recommendations are given concerning the maintenance, containment and transport of transgenic animals which are susceptible to pathogenic human viruses, with special attention to transgenic mice susceptible to polioviruses.

1993-01-01

379

Genetic and Molecular Analysis of Transgenic Rice cv. Rojolele Expressing Lactoferrin  

Directory of Open Access Journals (Sweden)

Full Text Available In a previous study, human lactoferrin gene have introduced into Javanica rice cv. Rojolele by Agrobacterium-mediated transformation. Lactoferrin (LF is an 80 kDa iron-binding glycoprotein that has been proposed to have many biological roles such as protection against microbial and virus infection. This study aims to analyze the integration and level of lactoferrin gene expression of transgenic rice cv. Rojolele. The study also aims to examine the genetic character of transgenic rice expressing recombinant lactoferrin. Stability expression of recombinant lactoferrin transgenic rice seeds over generations were analyzed by ELISA, while the integration stability of recombinant hLF gene in transgenic plants performed by PCR. The mitotic time, cell cycle and chromosome characterization of transgenic and non-transgenic rice cv. Rojolele were determined. Chromosome characterization of the trangenic and non transgenic rice cv. Rojolele was investigated to determine the genetic variation. All of the above efforts were aimed to evaluate the genetically engineered rice containing recombinant lactoferrin as a nutraceutical food. The results showed that the expression was stable through three consecutive generations. The expression of the hLF gene increased during grain-filling period. The active time of mitotic cells of transgenic rice rojolele was longer than the cells of non-transgenic rice. In addition, the cycle cell of transgenic and non-transgenic rojolele contained prophase, prometaphase, metaphase, anaphase, telophase and interphase. The result showed that all of the transgenic lines had diploid (2n chromosome number = 24.

Diah Rachmawati

2014-02-01

380

FACS purification of immunolabeled cell types from adult rat brain  

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Molecular analysis of brain tissue is greatly complicated by having many different classes of neurons and glia interspersed throughout the brain. Fluorescence-activated cell sorting (FACS) has been used to purify selected cell types from brain tissue. However, its use has been limited to brain tissue from embryos or transgenic mice with promoter-driven reporter genes. To overcome these limitations, we developed a FACS procedure for dissociating intact cell bodies from adult wild-type rat brai...

Guez-barber, Danielle; Fanous, Sanya; Harvey, Brandon K.; Zhang, Yongqing; Lehrmann, Elin; Becker, Kevin G.; Picciotto, Marina R.; Hope, Bruce T.

2012-01-01

 
 
 
 
381

In vivo mutation analysis using the ?X174 transgenic mouse and comparisons with other transgenes and endogenous genes.  

Science.gov (United States)

The ?X174 transgenic mouse was first developed as an in vivo Ames test, detecting base pair substitution (bps) at a single bp in a reversion assay. A forward mutational assay was also developed, which is a gain of function assay that also detects bps exclusively. Later work with both assays focused on establishing that a mutation was fixed in vivo using single-burst analysis: determining the number of mutant progeny virus from an electroporated cell by dividing the culture into aliquots before scoring mutants. We review results obtained from single-burst analysis, including testing the hypothesis that high mutant frequencies (MFs) of G:C to A:T mutation recovered by transgenic targets include significant numbers of unrepaired G:T mismatches. Comparison between the ?X174 and lacI transgenes in mouse spleen indicates that the spontaneous bps mutation frequency per nucleotide (mf(n)) is not significantly lower for ?X174 than for lacI; the response to ENU is also comparable. For the lacI transgene, the spontaneous bps mf(n) is highly age-dependent up to 12 weeks of age and the linear trend extrapolates at conception to a frequency close to the human bps mf(n) per generation of 1.7 × 10(-8). Unexpectedly, we found that the lacI somatic (spleen) bps mf(n) per cell division at early ages was estimated to be the same as for the human germ-line. The bps mf(n) in bone marrow for the gpt transgene is comparable to spleen for the lacI and ?X174 transgenes. We conclude that the G:C to A:T transition is characteristic of spontaneous in vivo mutation and that the MFs measured in these transgenes at early ages reflect the expected accumulation of in vivo mutation typical of endogenous mammalian mutation rates. However, spontaneous and induced mf(n)s per nucleotide for the cII gene in spleen are 5-10 times higher than for these other transgenes. PMID:20637298

Valentine, Carrie R; Delongchamp, Robert R; Pearce, Mason G; Rainey, Heather F; Dobrovolsky, Vasily N; Malling, Heinrich V; Heflich, Robert H

2010-12-01

382

Virus-Resistant Transgenic Plants Expressing L3.  

Science.gov (United States)

Disclosed are transgenic plants containing an exogenous nucleic acid encoding an L3 protein. The plant exhibits increased resistance to viruses and/or fungi that infect plants. The L3 proteins include wild-type proteins, spontaneously occurring mutants an...

J. D. Dinman K. A. Hudak N. E. Turner

2003-01-01

383

Efficient expression of transgenes in adult zebrafish by electroporation  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Expression of transgenes in muscle by injection of naked DNA is widely practiced. Application of electrical pulses at the site of injection was demonstrated to improve transgene expression in muscle tissue. Zebrafish is a precious model to investigate developmental biology in vertebrates. In this study we investigated the effect of electroporation on expression of transgenes in 3–6 month old adult zebrafish. Results Electroporation parameters such as number of pulses, voltage and amount of plasmid DNA were optimized and it was found that 6 pulses of 40 V·cm-1 at 15 ?g of plasmid DNA per fish increased the luciferase expression 10-fold compared to controls. Similar enhancement in transgene expression was also observed in Indian carp (Labeo rohita. To establish the utility of adult zebrafish as a system for transient transfections, the strength of the promoters was compared in A2 cells and adult zebrafish after electroporation. The relative strengths of the promoters were found to be similar in cell lines and in adult zebrafish. GFP fluorescence in tissues after electroporation was also studied by fluorescence microscopy. Conclusion Electroporation after DNA injection enhances gene expression 10-fold in adult zebrafish. Electroporation parameters for optimum transfection of adult zebrafish with tweezer type electrode were presented. Enhanced reporter gene expression upon electroporation allowed comparison of strengths of the promoters in vivo in zebrafish.

Rao S Hari

2005-10-01

384

SERPINB3 is associated with longer survival in transgenic mice.  

Science.gov (United States)

The physiological roles of the protease inhibitor SERPINB3 (SB3) are still largely unknown. The study was addressed to assess the biological effects of this serpin in vivo using a SB3 transgenic mouse model. Two colonies of mice (123 transgenic for SB3 and 148 C57BL/6J controls) have been studied. Transgenic (TG) mice showed longer survival than controls and the difference was more remarkable in males than in females (18.5% vs 12.7% life span increase). In TG mice decreased IL-6 in serum and lower p66shc in the liver were observed. In addition, TG males showed higher expression of mTOR in the liver. Liver histology showed age-dependent increase of steatosis and decrease of glycogen storage in both groups and none of the animals developed neoplastic lesions. In conclusion, the gain in life span observed in SB3-transgenic mice could be determined by multiple mechanisms, including the decrease of circulating IL-6 and the modulation of ageing genes in the liver. PMID:24162160

Villano, Gianmarco; Ruvoletto, Mariagrazia; Ceolotto, Giulio; Quarta, Santina; Calabrese, Fiorella; Turato, Cristian; Tono, Natascia; Biasiolo, Alessandra; Cattelan, Arianna; Merkel, Carlo; Avogaro, Angelo; Gatta, Angelo; Pontisso, Patrizia

2013-01-01

385

HER-2/neu x Aromatase Double Transgenic Mice Model  

Science.gov (United States)

A majority of breast cancers are hormone-responsive, and require estrogen for growth, and respond to hormonal therapy that blocks estrogen receptor action. Breast tumors with low levels of or completely lacking estrogen receptor fail to respond to antiestrogen therapy yet require estrogen for tumor initiation. To address the importance of local estrogen in oncogene-mediated breast tumorigenesis, we have crossed MMTV-aromatase with MMTV-HER2/neu and examined the incidence of breast cancer in double transgenic mice in comparison with parental strains. Double transgenic mice show normal mammary development and express both transgenes at similar levels to that of parental strains. Tumor incidence in double transgenic mice ( 65%). In addition to a significant decrease in tumorigenesis, these mice expressed ER? as well as high levels of ER? along with decreased levels of cyclin D1 and phosphorylated pRb among other changes. Furthermore, experiments using THC (ER?- agonist and ER?-antagonist) clearly demonstrate the critical role of ER? in HER2/neu-mediated tumorigenesis. These studies provide the first genetic evidence that estrogen receptor, mainly ER? than ER? and its dependent changes play an important role in regulating mammary tumorigenesis. These findings provide further evidence for development and testing of novel therapeutic approaches based on selective regulation of estrogen receptors (ER? and ?) - dependent actions for the treatment and prevention of breast cancers.

Tekmal, Rajeshwar Rao; Nair, Hareesh B.; Perla, Rao P.; Kirma, Nameer

2007-01-01

386

Transgenic plastids in basic research and plant biotechnology.  

Science.gov (United States)

Facile methods of genetic transformation are of outstanding importance for both basic and applied research. For many years, transgenic technologies for plants were restricted to manipulations of the nuclear genome. More recently, a second genome of the plant cell has become amenable to genetic engineering: the prokaryotically organized circular genome of the chloroplast. The possibility to directly manipulate chloroplast genome-encoded information has paved the way to detailed in vivo studies of virtually all aspects of plastid gene expression. Moreover, plastid transformation technologies have been intensely used in functional genomics by performing gene knockouts and site-directed mutageneses of plastid genes. These studies have contributed greatly to our understanding of the physiology and biochemistry of biogenergetic processes inside the plastid compartment. Plastid transformation technologies have also stirred considerable excitement among plant biotechnologists, since transgene expression from the plastid genome offers a number of most attractive advantages, including high-level foreign protein expression and transgene containment due to lack of pollen transmission. This review describes the generation of plants with transgenic plastids, summarizes our current understanding of the transformation process and highlights selected applications of transplastomic technologies in basic and applied research. PMID:11563907

Bock, R

2001-09-21

387

APPLICATION OF SPECTRAL IMAGING TO TRANSGENIC CORN MONITORING  

Science.gov (United States)

Transgenic crops containing pesticidal traits are regulated by EPA under the Federal Insecticide Fungicide and Rodenticide Act. The EPA has declared crops engineered to contain a bacterial gene from Bacillus thuringiensis (Bt) to be in the public good, due to their potential to...

388

Visualization of C. elegans transgenic arrays by GFP  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Targeting the green fluorescent protein (GFP via the E. coli lac repressor (LacI to a specific DNA sequence, the lac operator (lacO, allows visualization of chromosomes in yeast and mammalian cells. In principle this method of visualization could be used for genetic mosaic analysis, which requires cell-autonomous markers that can be scored easily and at single cell resolution. The C. elegans lin-3 gene encodes an epidermal growth factor family (EGF growth factor. lin-3 is expressed in the gonadal anchor cell and acts through LET-23 (transmembrane protein tyrosine kinase and ortholog of EGF receptor to signal the vulval precursor cells to generate vulval tissue. lin-3 is expressed in the vulval cells later, and recent evidence raises the possibility that lin-3 acts in the vulval cells as a relay signal during vulval induction. It is thus of interest to test the site of action of lin-3 by mosaic analysis. Results We visualized transgenes in living C. elegans by targeting the green fluorescent protein (GFP via the E. coli lac repressor (LacI to a specific 256 sequence repeat of the lac operator (lacO incorporated into transgenes. We engineered animals to express a nuclear-localized GFP-LacI fusion protein. C. elegans cells having a lacO transgene result in nuclear-localized bright spots (i.e., GFP-LacI bound to lacO. Cells with diffuse nuclear fluorescence correspond to unbound nuclear localized GFP-LacI. We detected chromosomes in living animals by chromosomally integrating the array of the lacO repeat sequence and visualizing the integrated transgene with GFP-LacI. This detection system can be applied to determine polyploidy as well as investigating chromosome segregation. To assess the GFP-LacI•lacO system as a marker for mosaic analysis, we conducted genetic mosaic analysis of the epidermal growth factor lin-3, expressed in the anchor cell. We establish that lin-3 acts in the anchor cell to induce vulva development, demonstrating this method's utility in detecting the presence of a transgene. Conclusion The GFP-LacI•lacO transgene detection system works in C. elegans for visualization of chromosomes and extrachromosomal transgenes. It can be used as a marker for genetic mosaic analysis. The lacO repeat sequence as an extrachromosomal array becomes a valuable technique allowing rapid, accurate determination of spontaneous loss of the array, thereby allowing high-resolution mosaic analysis. The lin-3 gene is required in the anchor cell to induce the epidermal vulval precursors cells to undergo vulval development.

Sternberg Paul W

2006-06-01

389

Activation of transgene expression by early region 4 is responsible for a high level of persistent transgene expression from adenovirus vectors in vivo.  

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The persistence of transgene expression has become a hallmark for adenovirus vector evaluation in vivo. Although not all therapeutic benefit in gene therapy is reliant on long-term transgene expression, it is assumed that the treatment of chronic diseases will require significant persistence of expression. To understand the mechanisms involved in transgene persistence, a number of adenovirus vectors were evaluated in vivo in different strains of mice. Interestingly, the rate of vector genome ...

1997-01-01

390

Development and utilization of transgenic New World screwworm, Cochliomyia hominivorax.  

Science.gov (United States)

The New World screwworm (NWS), Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae), was the first insect to be effectively controlled using the sterile insect technique (SIT). Recent efforts to improve SIT control of this species have centred on the development of genetically transformed strains using the piggyBac transposon vector system. Eight transgenic strains were produced incorporating an enhanced green fluorescent protein (EGFP) marker gene under polyubiquitin regulation that has the potential for use as a genetic marking system for released males. The tr