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1

A monoclonal antibody against kininogen reduces inflammation in the HLA-B27 transgenic rat  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The human leukocyte antigen B27 (HLA-B27) transgenic rat is a model of human inflammatory bowel disease, rheumatoid arthritis and psoriasis. Studies of chronic inflammation in other rat models have demonstrated activation of the kallikrein–kinin system as well as modulation by a plasma kallikrein in...

Keith, James C; Sainz, Irma M; Isordia-Salas, Irma; Pixley, Robin A; Leathurby, Yelena; Albert, Leo M; Colman, Robert W

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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 c...

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

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HLA-B27 misfolding and ankylosing spondylitis.  

UK PubMed Central (United Kingdom)

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.

Colbert RA; Tran TM; Layh-Schmitt G

2014-01-01

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

MHC class I allele, HLA-B27, is strongly associated with a group of human diseases called spondyloarthropathies. Some of these diseases have an onset after an enteric or genitourinary infection. In the present study, we describe spontaneous disease in HLA-B27 transgenic mice where endogenous beta2-m...

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

5

HLA-B27 associated arthropathies.  

UK PubMed Central (United Kingdom)

The association between histo-compatibility antigens and disease is reviewed, in particular that between HLA-B27 and spondylitic disorders, i.e., ankylosing spondylitis, Reiter's arthritis, psoriatic arthritis, and ankylosing hyperostosis. We determined whether the presence of HLA-B27 predicted specific radiographic findings and, conversely, whether specific radiographic changes predicted antigenic status. The prevalences of the HLA-B27 antigen in our patients were: ankylosing spondylitis, 100%; Reiter's arthritis, 93%; psoriatic arthritis, 55%; and ankylosing hyperostosis, 12%. The only specific radiographic finding associated with B27 positivity was severe spondylitis in psoriasis.

Schumacher TM; Genant HK; Kellet MJ; Mall JC; Fye KH

1978-02-01

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HLA-B27 associated arthropathies.  

Science.gov (United States)

The association between histo-compatibility antigens and disease is reviewed, in particular that between HLA-B27 and spondylitic disorders, i.e., ankylosing spondylitis, Reiter's arthritis, psoriatic arthritis, and ankylosing hyperostosis. We determined whether the presence of HLA-B27 predicted specific radiographic findings and, conversely, whether specific radiographic changes predicted antigenic status. The prevalences of the HLA-B27 antigen in our patients were: ankylosing spondylitis, 100%; Reiter's arthritis, 93%; psoriatic arthritis, 55%; and ankylosing hyperostosis, 12%. The only specific radiographic finding associated with B27 positivity was severe spondylitis in psoriasis. PMID:622470

Schumacher, T M; Genant, H K; Kellet, M J; Mall, J C; Fye, K H

1978-02-01

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Polymorphism of HLA-B27: 105 Subtypes Currently Known.  

Science.gov (United States)

HLA-B27 has a high degree of genetic polymorphism, with 105 known subtypes, named HLA-B*27:01 to HLA-B*27:106, encoded by 132 alleles. The most common subtypes associated with ankylosing spondylitis are HLA-B*27:05 (Caucasians), HLA-B*27:04 (Chinese), and HLA-B*27:02 (Mediterranean populations). For Chinese populations, HLA-B*27:04 is associated with a greater ankylosing spondylitis risk than HLA-B*27:05. Two subtypes, HLA-B27*06 and HLA-B27*09, seem to have no disease association. These differential disease associations of HLA-B27 subtypes, and the recent discovery that ERAP1 is associated with ankylosing spondylitis for patients with HLA-B27, have increased attempts to determine the function of HLA-B27 in disease pathogenesis by studying hemodynamic features of its protein structure, alterations of its peptidome, aberrant peptide handling, and associated molecular events. However, after 40 years we still do not fully know how HLA-B27 predisposes to ankylosing spondylitis and related spondyloarthritis. PMID:23990399

Khan, Muhammad Asim

2013-10-01

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Polymorphism of HLA-B27: 105 Subtypes Currently Known.  

UK PubMed Central (United Kingdom)

HLA-B27 has a high degree of genetic polymorphism, with 105 known subtypes, named HLA-B*27:01 to HLA-B*27:106, encoded by 132 alleles. The most common subtypes associated with ankylosing spondylitis are HLA-B*27:05 (Caucasians), HLA-B*27:04 (Chinese), and HLA-B*27:02 (Mediterranean populations). For Chinese populations, HLA-B*27:04 is associated with a greater ankylosing spondylitis risk than HLA-B*27:05. Two subtypes, HLA-B27*06 and HLA-B27*09, seem to have no disease association. These differential disease associations of HLA-B27 subtypes, and the recent discovery that ERAP1 is associated with ankylosing spondylitis for patients with HLA-B27, have increased attempts to determine the function of HLA-B27 in disease pathogenesis by studying hemodynamic features of its protein structure, alterations of its peptidome, aberrant peptide handling, and associated molecular events. However, after 40 years we still do not fully know how HLA-B27 predisposes to ankylosing spondylitis and related spondyloarthritis.

Khan MA

2013-10-01

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HLA B27 y las espondilartropatías seronegativas  

Directory of Open Access Journals (Sweden)

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

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

1997-01-01

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HLA-B27 subtypes among the Chukotka native groups  

International Nuclear Information System (INIS)

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

1995-01-01

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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 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 com 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 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.

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

2008-10-01

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

Directory of Open Access Journals (Sweden)

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

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

2008-01-01

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Improved flow cytometric method for HLA-B27 typing.  

Science.gov (United States)

HLA-B27 is a cell marker of clinical interest because of its high association with certain diseases. The HLA-B27 antigen was detected on lymphocytes using a monoclonal antibody in an indirect immuno-fluorescence assay using a fluorescence flow cytometer. The considerable crossreaction of the monoclonal antibody with the HLA-B7 antigen was effectively suppressed by masking it by means of human anti-HLA-B7 antiserum. The flow cytometric method was evaluated by comparing the results with those obtained by the standard lymphocytotoxicity test and showed complete agreement in 107 selected patient samples. PMID:1489163

Janssen, W C; Rouwen, J A; Hoffmann, J J

1992-11-01

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Improved flow cytometric method for HLA-B27 typing.  

UK PubMed Central (United Kingdom)

HLA-B27 is a cell marker of clinical interest because of its high association with certain diseases. The HLA-B27 antigen was detected on lymphocytes using a monoclonal antibody in an indirect immuno-fluorescence assay using a fluorescence flow cytometer. The considerable crossreaction of the monoclonal antibody with the HLA-B7 antigen was effectively suppressed by masking it by means of human anti-HLA-B7 antiserum. The flow cytometric method was evaluated by comparing the results with those obtained by the standard lymphocytotoxicity test and showed complete agreement in 107 selected patient samples.

Janssen WC; Rouwen JA; Hoffmann JJ

1992-11-01

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Psoriatic spondyloarthropathy: a comparative study between HLA-B27 positive and HLA-B27 negative disease.  

UK PubMed Central (United Kingdom)

OBJECTIVES: To evaluate the relative contribution of the human leukocyte antigen (HLA)-B27 to psoriatic spondyloarthropathy (PsSpA) susceptibility and to analyze whether this antigen contributes to disease expression. METHODS: This cross-sectional study included 70 patients (mean age 48 +/- 14.5 years; 44 men and 26 women). PsSpA was defined according to radiological findings (grade 2 or more sacroiliitis), and patients were classified into 3 main subtypes: isolated axial disease (n = 16), axial plus oligoarthritis (n = 29) and axial plus polyarthritis (n = 25). All patients were studied following a standard protocol that included the collection of demographic and epidemiological data, clinical history, radiographs, complementary tests, physical examination, and HLA-B27 testing (serological method). For functional evaluation, the Health Assessment Questionnaire-Specific for spondyloarthropathy (HAQ-S) was used. Patients with and without HLA-B27 antigen were compared on the basis of the data. RESULTS: Twenty-four patients (34%) carried the HLA-B27 antigen (RR 6.4, P <.0004). Fifty-six percent of those patients with the isolated axial pattern had this antigen, compared with 24% in the poly-arthritis axial pattern and 31% of those in the oligo-arthritis axial group (P =.016). Univariate analysis demonstrated correlations between HLA-B27 and an earlier age of onset for both psoriasis (P =.028) and arthritis (P =.006), male gender (P =.002), bilateral sacroiliitis (P =.002), and uveitis (P =.026). HLA-B27 negative patients developed more peripheral erosions than HLA-B27 positive patients (P =.05). No correlation was found between B27 and clinical symptoms of back involvement, syndesmophytes, or functional impairment. CONCLUSIONS: The HLA-B27 antigen is not only important for PsSpA susceptibility, but also determines some clinical features. This antigen was associated with earlier age of psoriasis and arthritis onset, bilateral sacroiliitis, and male gender. However, it was not associated with either the severity or extension of the spondylitic process or with functional impairment.

Queiro R; Sarasqueta C; Belzunegui J; Gonzalez C; Figueroa M; Torre-Alonso JC

2002-06-01

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

UK PubMed Central (United Kingdom)

Objectives. HLA-B*27:05 is associated with AS whereas HLA-B*27:09 is not associated. We hypothesized that different interactions with KIR immune receptors could contribute to the difference in disease association between HLA-B*27:05 and HLAB*27:09. Thus, the objective of this study was to compare the formation of ?2m-free heavy chain (FHC) including B27 dimers (B272) by HLA-B*27:05 and HLA-B*27:09 and their binding to KIR immunoreceptors.Methods. We studied the formation of HLA-B*27:05 and HLA-B*27:09 heterotrimers and FHC forms including dimers in vitro and in transfected cells. We investigated HLA-B*27:05 and HLA-B*27:09 binding to KIR3DL1, KIR3DL2 and LILRB2 by FACS staining with class I tetramers and by quantifying interactions with KIR3DL2CD3?-reporter cells and KIR3DL2-expressing NK cells. We also measured KIR expression on peripheral blood NK and CD4 T cells from 18 HLA-B*27:05 AS patients, 8 HLA-B27 negative and 12 HLA-B*27:05+ and HLA-B*27:09+ healthy controls by FACS staining.Results. HLA-B*27:09 formed less B272 and FHC than HLA-B*27:05. HLA-B*27:05-expressing cells stimulated KIR3DL2CD3?-reporter T cells more effectively. Cells expressing HLA-B*27:05 promoted KIR3DL2+ NK cell survival more strongly than HLA-B*27:09. HLA-B*27:05 and HLA-B*27:09 dimer tetramers stained KIR3DL1, KIR3DL2 and LILRB2 equivalently. Increased proportions of NK and CD4 T cells expressed KIR3DL2 in HLA-B*27:05+ AS patients compared with HLA-B*27:05+, HLA-B*27:09+ and HLA-B27- healthy controls.Conclusion. Differences in the formation of FHC ligands for KIR3DL2 by HLA-B*27:05 and HLA-B*27:09 could contribute to the differential association of these alleles with AS.

Cauli A; Shaw J; Giles J; Hatano H; Rysnik O; Payeli S; McHugh K; Dessole G; Porru G; Desogus E; Fiedler S; Hölper S; Carette A; Blanco-Gelaz MA; Vacca A; Piga M; Ibba V; Garau P; La Nasa G; López-Larrea C; Mathieu A; Renner C; Bowness P; Kollnberger S

2013-06-01

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HLA-B27 and clinical features of acute anterior uveitis in Cuba.  

UK PubMed Central (United Kingdom)

BACKGROUND: There are few data about the epidemiology of acute anterior uveitis (AAU) from Latin America. In Cuba, the genetic admixture of the population could modify the HLA-B27-AAU association. In this study, the authors compared the distribution of the HLA-B27 allele in patients and controls and described some clinical outcomes. MATERIALS AND METHODS: The clinical features of patients were collected from their medical records. HLA-B27 genotyping was performed using the polymerase chain reaction. HLA-B27 allele distribution was compared between patients and controls. RESULTS: HLA-B27 allele was present in 55.4% of the patients and 0.87% of the controls. AAU HLA-B27 positivity was associated with males, frequent episodes, and a systemic disease. There is no difference in ocular complications between HLA-B27-positive and -negative patients. CONCLUSIONS: Results from this study are similar to data described in other countries. HLA-B27 allele distribution in controls is lower than other reports in Caucasian populations.

Torres S; Borges S; Artiles A

2013-04-01

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The pathogenesis of HLA-B27 associated arthritis: lessons from the B27 crystal.  

UK PubMed Central (United Kingdom)

The most remarkable association between a major histocompatibility complex antigen and disease susceptibility--HLA-B27 and seronegative spondyloarthropathies, particularly ankylosing spondylitis--was discovered 20 years ago. During the two intervening decades advances in basic immunology and molecular biology have not only revealed the biosynthesis and structure of HLA-B27 but also given clues to the basic function of this molecule, the presentation of allele-specific peptides to CD8+ cytotoxic T cells. The recently reported three-dimensional structure of HLA-B27 and the identification of self-peptides bound to this major histocompatibility complex class I antigen can be viewed as a landmark in the understanding of the pathogenic role of HLA-B27. Based on crystallographic evidence, a peptide-binding motif can be postulated that should allow identification of HLA-B27 complexed peptides which may trigger an immune reaction causing arthritis.

Kellner H; Yu D

1994-03-01

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

Directory of Open Access Journals (Sweden)

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

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

2012-01-01

20

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

Scientific Electronic Library Online (English)

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

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

2012-12-01

 
 
 
 
21

HLA-B27 frequency in a group of patients with psoriatic arthritis*  

Science.gov (United States)

BACKGROUND HLA-B27 is associated with spondyloarthritis, a group of diseases that includes psoriatic arthritis. OBJECTIVES To describe the HLA-B27 frequency in a group of Brazilian patients with psoriatic arthritis and correlate its presence or absence with their clinical manifestations. METHODS Cross-sectional study with 44 psoriatic arthritis patients of a Rheumatology clinic. Demographic and social data were recorded, as were skin and joints clinical examination. HLA-B27 was tested. All data were processed descriptively and comparatively by appropriate software. Parametric and non parametric tests were used with 5% statistical significance. RESULTS HLA-B27 was negative in 32 of the 44 patients (72,7%). Most of them were male, Caucasian, living in Rio de Janeiro, with plaque type psoriasis and average age of 52,9 years. There was statistical significant correlation between positive HLA-B27 and male gender (p=0,004). Negative HLA-B27 had a tendency to correlate with hands and wrists arthritis (p=0,07). There was an inverse significant correlation between HLA values and Schöber's test (p=0,02). CONCLUSION Although HLA-B27 is negative in most of patients, it is significantly associated to male gender and inversely correlated with Schöber's test.

Ruiz, Danilo Garcia; de Azevedo, Mario Newton Leitao; Lupi, Omar

2012-01-01

22

HLA-B27 frequency in a group of patients with psoriatic arthritis.  

UK PubMed Central (United Kingdom)

BACKGROUND: HLA-B27 is associated with spondyloarthritis, a group of diseases that includes psoriatic arthritis. OBJECTIVES: To describe the HLA-B27 frequency in a group of Brazilian patients with psoriatic arthritis and correlate its presence or absence with their clinical manifestations. METHODS: Cross-sectional study with 44 psoriatic arthritis patients of a Rheumatology clinic. Demographic and social data were recorded, as were skin and joints clinical examination. HLA-B27 was tested. All data were processed descriptively and comparatively by appropriate software. Parametric and non parametric tests were used with 5% statistical significance. RESULTS: HLA-B27 was negative in 32 of the 44 patients (72,7%). Most of them were male, Caucasian, living in Rio de Janeiro, with plaque type psoriasis and average age of 52,9 years. There was statistical significant correlation between positive HLA-B27 and male gender (p=0,004). Negative HLA-B27 had a tendency to correlate with hands and wrists arthritis (p=0,07). There was an inverse significant correlation between HLA values and Schöber's test (p=0,02). CONCLUSION: Although HLA-B27 is negative in most of patients, it is significantly associated to male gender and inversely correlated with Schöber's test.

Ruiz DG; Azevedo MN; Lupi O

2012-11-01

23

Clinical Analysis of 240 Patients with HLA-B27 Associated Acute Anterior Uveitis.  

UK PubMed Central (United Kingdom)

PURPOSE: To analyze the prevalence of HLA-B27 associated acute anterior uveitis and to identify its clinical features. METHODS: A total of 240 patients with HLA-B27 associated acute anterior uveitis, who were admitted to Zhejiang Ophthalmologic Hospital between December 2006 and October 2012, were retrospectively analyzed. The age of onset, sex, affected eyes, HLA-B27 antigen detection, recurrence, joint involvement, and surgical complications were investigated. RESULTS: The average age of onset was 37.0±12.0 years and the ratio of male to female patients was 2.4:1. Most cases had alternate unilateral or bilateral involvement. Among all participants, 234 cases (97.5%) were HLA-B27 positive, and 124 cases (51.7%) had spondyloarthropathies (SpA), dominated by 108 cases with ankylosing spondylitis (AS,45.0%), and mostly seen in male subjects (P<0.05). Six patients were HLA-B27 negative (2.5%) and no statistical significance was noted between male and female patients (P>0.05). A total of 193 cases (80.4%) presented with complications, mainly fibrinous exudation, posterior synechia, and vitreous opacity. CONCLUSION: HLA-B27 that is associated acute anterior uveitis with a relatively high incidence and recurrence presents with more severe clinical features than does idiopathic acute anterior uveitis, and it often accompanies systemic arthritic diseases. HLA-B27 antibody detection is associated with the diagnosis and treatment of acute anterior uveitis.

Zheng MQ; Wang YQ; Lu XY; Wang YL; Mao LP; Gu YF; Chen PF

2012-12-01

24

Immunoradiometric assay for the rapid detection of HLA-B27.  

UK PubMed Central (United Kingdom)

A new method for the rapid detection of the HLA-B27 antigen is discussed which consists of a direct immunoradiometric assay (IRA) utilizing a 125I-labelled, HLA-B27 specific monoclonal antibody to detect HLA-B27 in whole citrated blood. Thus far, the assay has been used to assign HLA-B27 status in a population study involving 142 individuals; 36 subjects were HLA-B27+ by conventional microcytotoxicity and all 36 were detected by the new IRA and all 106 B27- individuals were also accurately assigned by IRA. The problem of detection of the cross-reactive HLA antigen HLA-B7 was overcome by blocking the reactive sites on HLA-B7 molecules with the HLA-B7 specific antibody, BB7.1. This new assay enables accurate detection of HLA-B27 within 30 min of venepuncture and should facilitate the assessment of HLA-B27 status in tissue typing laboratories.

Trapani JA; Vaughan HA; Tait BD; McKenzie IF

1988-01-01

25

Immunoradiometric assay for the rapid detection of HLA-B27.  

Science.gov (United States)

A new method for the rapid detection of the HLA-B27 antigen is discussed which consists of a direct immunoradiometric assay (IRA) utilizing a 125I-labelled, HLA-B27 specific monoclonal antibody to detect HLA-B27 in whole citrated blood. Thus far, the assay has been used to assign HLA-B27 status in a population study involving 142 individuals; 36 subjects were HLA-B27+ by conventional microcytotoxicity and all 36 were detected by the new IRA and all 106 B27- individuals were also accurately assigned by IRA. The problem of detection of the cross-reactive HLA antigen HLA-B7 was overcome by blocking the reactive sites on HLA-B7 molecules with the HLA-B7 specific antibody, BB7.1. This new assay enables accurate detection of HLA-B27 within 30 min of venepuncture and should facilitate the assessment of HLA-B27 status in tissue typing laboratories. PMID:3155158

Trapani, J A; Vaughan, H A; Tait, B D; McKenzie, I F

1988-01-01

26

Interferon-? contributes to HLA-B27-associated unfolded protein response in spondyloarthropathies.  

UK PubMed Central (United Kingdom)

OBJECTIVE: HLA-B27 positivity strongly influences the susceptibility to and phenotype of spondyloarthropathies (SpA). This study was designed to screen factors that activate the promoter of HLA-B27 in U937 cells, and to assess whether these promoter-activating factors induce the unfolded protein response (UPR) in HLA-B27-expressing cells. METHODS: Cytometric Bead Array, flow cytometry, and real-time polymerase chain reaction were used to detect the expression of cytokines and UPR-associated proteins in peripheral blood and synovial fluid of patients with SpA. The HLA-B27 promotor transfectant was incubated separately with cytokines and Toll-like receptor ligands. After interferon-? (IFN-?) stimulation, expressions of GRP78, CHOP, and XBP-1 were tested in HLA-B27-expressing U937 cells and peripheral blood mononuclear cell (PBMC) of patients with ankylosing spondylitis (AS). (Clinical trial registration no. ChiCTR-OCC-11001565) RESULTS: Expressions of GRP78, CHOP, and XBP-1 in monocytes/macrophages of SpA peripheral blood and synovial fluid were higher than those in healthy controls and patients with osteoarthritis (OA) (p < 0.05). Tumor necrosis factor-? (TNF-?) and IFN-?, IFN-ß, and IFN-? were found to have activated the HLA-B27 promoter in the U937 cell line (p < 0.05). Following stimulation with IFN-?, the expressions of GRP78, CHOP and XBP-1 in HLA-B27-transfected U937 cells and PBMC of HLA-B27-positive AS patients were more intense than those in A2-U937 cells, HLA-B27-negative AS patients, or healthy controls (p < 0.05). CONCLUSION: Expressions of GRP78, CHOP, and XBP-1 were higher in monocytes/macrophages of patients with SpA than those in both OA patients and healthy controls, suggesting that UPR may participate in the pathogenesis of SpA. TNF-? and IFN-?, IFN-ß, and IFN-? significantly activated HLA-B27 promoter in the U937 cell line, and IFN-?, the strongest activating factor, may induce the UPR in HLA-B27-expressing cells.

Feng Y; Ding J; Fan CM; Zhu P

2012-03-01

27

Prevalence of HLA-B27 in Patients with Ankylosing Spondylitis in Qatar.  

Science.gov (United States)

Background and Objectives. The human leukocyte antigen HLA-B27 is a class 1 antigen of the major histocompatibility complex and is strongly associated with ankylosing spondylitis (AS). The purpose of the present study is to investigate the distribution of HLA-B27 in patients with AS of different ethnic groups in Qatar. Design and Setting. Study design was cross-sectional and the setting was rheumatology clinics of Hamad General Hospital in Qatar where most of ankylosing spondylitis patients are followed up. Patients and Methods. Patients with diagnosis of AS who met the New York modified criteria for AS were tested for HLA-B27. 119 patients were tested for HLA-B27: 66 Arabs, 52 Asians (Indians, Pakistanis, Bengalis, and Iranians), and one Western (Irish). Results. Of all the individuals, 82 were positive (69%) for HLA-B27. Among the Arabs, 49/66 were positive (74%). Among the Asians, 32/52 were positive (61%). Furthermore, Qatari patients (10 males and one female) 9 were positive (82%), 14/19 Jordanians/Palestinians were positive, and 9/10 (90%) Egyptians were positive. Among the Asians, 19/26 Indians were positive (73%), which was similar to the Arabs. Conclusion. HLA-B27 in our small group of Arabs is present in 74%. Comparison with other data will be presented in detail. PMID:22548073

Abdelrahman, M H; Mahdy, S; Khanjar, I A; Siam, A M; Malallah, H A; Al-Emadi, S A; Sarakbi, H A; Hammoudeh, M

2012-04-04

28

Prevalence of HLA-B27 in Patients with Ankylosing Spondylitis in Qatar.  

UK PubMed Central (United Kingdom)

Background and Objectives. The human leukocyte antigen HLA-B27 is a class 1 antigen of the major histocompatibility complex and is strongly associated with ankylosing spondylitis (AS). The purpose of the present study is to investigate the distribution of HLA-B27 in patients with AS of different ethnic groups in Qatar. Design and Setting. Study design was cross-sectional and the setting was rheumatology clinics of Hamad General Hospital in Qatar where most of ankylosing spondylitis patients are followed up. Patients and Methods. Patients with diagnosis of AS who met the New York modified criteria for AS were tested for HLA-B27. 119 patients were tested for HLA-B27: 66 Arabs, 52 Asians (Indians, Pakistanis, Bengalis, and Iranians), and one Western (Irish). Results. Of all the individuals, 82 were positive (69%) for HLA-B27. Among the Arabs, 49/66 were positive (74%). Among the Asians, 32/52 were positive (61%). Furthermore, Qatari patients (10 males and one female) 9 were positive (82%), 14/19 Jordanians/Palestinians were positive, and 9/10 (90%) Egyptians were positive. Among the Asians, 19/26 Indians were positive (73%), which was similar to the Arabs. Conclusion. HLA-B27 in our small group of Arabs is present in 74%. Comparison with other data will be presented in detail.

Abdelrahman MH; Mahdy S; Khanjar IA; Siam AM; Malallah HA; Al-Emadi SA; Sarakbi HA; Hammoudeh M

2012-01-01

29

Association of HLA B27 antigen in Indian patients of ankylosing spondylitis and other autoimmune diseases.  

UK PubMed Central (United Kingdom)

One thousand three hundred and forty clinically suspected patients of Ankylosing Spondylitis (AS) and other autoimmune diseases and 5000 controls were studied to detect the association of HLA B27 antigen amongst them. Other alleles studied include HLA B7, B40 (B60), B22(B55), B13, etc. Our findings show a considerable and consistent association of HLA B27 with AS irrespective of the community to which the patient, belonged his hygiene or socio-economic conditions. We also found that people in the age group of 21-39 were the most vulnerable, when number of affected individuals or severity of the disease were taken into consideration. Male members showed a preponderance over females in HLA B27 positivity. Detection of HLA B27 could help in the diagnosis of AS. Patients suffering from other autoimmune diseases such as rheumatoid arthritis, psoriasis, Reiter's syndrome and uveitis and patients with inflammatory bowel disease, colitis, eczema, bacillary or fungal infection were also found to be HLA B27 positive. A study of other alleles shows that even they sometimes associate AS and other autoimmune diseases.

Kankonkar SR; Raikar SC; Joshi SV; Tijoriwala SJ

1998-04-01

30

Salivary endoscopy in a pediatric patient with HLA-B27 seropositivity and recurrent submandibular sialadenitis.  

Science.gov (United States)

Patients with human leukocyte antigen (HLA)-B27 seropositivity have a genetic predisposition to form spondyloarthropathies, especially ankylosing spondylitis. Other related inflammatory or autoimmune disorders include reactive arthritis, uveitis, psoriatic arthritis, and Crohn's disease. Although juvenile recurrent parotitis is not uncommon, recurrent submandibular sialadenitis is rare in pediatric patients. Sialadenitis is typically caused by salivary stones, infection, or duct stricture. To our knowledge, there has not been report of HLA-B27 positivity and recurrent sialadenitis described previously. We describe a patient with HLA-B27 seropositivity and multiple episodes of left submandibular sialadenitis who underwent diagnostic and therapeutic sialendoscopy. Previous treatment included antibiotics, sialogogues, warm compresses, and hydration before he underwent definitive sialendoscopy treatment at a tertiary care medical center. Salivary endoscopy showed salivary stasis and sludging within the left submandibular gland duct, with no salivary stones. Topical steroid was applied to the duct. At one year following his surgery, he has not had any recurrent episodes of sialadenitis. HLA-B27 seropositivity is associated with many inflammatory disorders; we report a case in which the patient had coexisting recurrent sialadenitis. In the pediatric population, sialadenitis is traditionally managed with antibiotics and supportive care, however our patient underwent salivary endoscopy. Sialendoscopy is an emerging modality that potentially avoids radiation exposure from CT or sialography and should be considered as another preferred treatment option. More investigation is required to prove a possible correlation between existing HLA-B27 and the propensity to develop this clinical problem. PMID:23639340

Nguyen, Amy M; Francis, Carrie L; Larsen, Christopher G

2013-04-29

31

Salivary endoscopy in a pediatric patient with HLA-B27 seropositivity and recurrent submandibular sialadenitis.  

UK PubMed Central (United Kingdom)

Patients with human leukocyte antigen (HLA)-B27 seropositivity have a genetic predisposition to form spondyloarthropathies, especially ankylosing spondylitis. Other related inflammatory or autoimmune disorders include reactive arthritis, uveitis, psoriatic arthritis, and Crohn's disease. Although juvenile recurrent parotitis is not uncommon, recurrent submandibular sialadenitis is rare in pediatric patients. Sialadenitis is typically caused by salivary stones, infection, or duct stricture. To our knowledge, there has not been report of HLA-B27 positivity and recurrent sialadenitis described previously. We describe a patient with HLA-B27 seropositivity and multiple episodes of left submandibular sialadenitis who underwent diagnostic and therapeutic sialendoscopy. Previous treatment included antibiotics, sialogogues, warm compresses, and hydration before he underwent definitive sialendoscopy treatment at a tertiary care medical center. Salivary endoscopy showed salivary stasis and sludging within the left submandibular gland duct, with no salivary stones. Topical steroid was applied to the duct. At one year following his surgery, he has not had any recurrent episodes of sialadenitis. HLA-B27 seropositivity is associated with many inflammatory disorders; we report a case in which the patient had coexisting recurrent sialadenitis. In the pediatric population, sialadenitis is traditionally managed with antibiotics and supportive care, however our patient underwent salivary endoscopy. Sialendoscopy is an emerging modality that potentially avoids radiation exposure from CT or sialography and should be considered as another preferred treatment option. More investigation is required to prove a possible correlation between existing HLA-B27 and the propensity to develop this clinical problem.

Nguyen AM; Francis CL; Larsen CG

2013-06-01

32

The radiographic features of rheumatoid arthritis in HLA-B27-positive patients  

International Nuclear Information System (INIS)

Radiographs were reviewed in a group of nine patients with classical seropositive rheumatoid arthritis who on tissue typing were found to express the class I HLA-B27 allele. Radiographs were analyzed with regard to whether or not they demonstrated radiographic features of (1) classical rheumatoid arthritis, (2) seronegative arthritis, or (3) mixed features of rheumatoid and seronegative arthritis. Five patients (55%) displayed radiographic features consistent with a diagnosis of rheumatoid arthritis, two patients (22%) showed radiographic features of seronegative disorder (periostitis and sacroiliitis), and two patients (22%) showed a mixed picture with evidence of both rheumatoid arthritis and a seronegative disorder. Thus, the HLA-B27 allele contributed to the radiographic features in 44% of patients with rheumatoid arthritis and associated HLA-B27. Thus, the wide range of findings in our population indicates that the radiographic attributes are not specific enough to constitute a unique subpopulation of patients with rheumatoid arthritis. (orig.)

1993-01-01

33

The radiographic features of rheumatoid arthritis in HLA-B27-positive patients  

Energy Technology Data Exchange (ETDEWEB)

Radiographs were reviewed in a group of nine patients with classical seropositive rheumatoid arthritis who on tissue typing were found to express the class I HLA-B27 allele. Radiographs were analyzed with regard to whether or not they demonstrated radiographic features of (1) classical rheumatoid arthritis, (2) seronegative arthritis, or (3) mixed features of rheumatoid and seronegative arthritis. Five patients (55%) displayed radiographic features consistent with a diagnosis of rheumatoid arthritis, two patients (22%) showed radiographic features of seronegative disorder (periostitis and sacroiliitis), and two patients (22%) showed a mixed picture with evidence of both rheumatoid arthritis and a seronegative disorder. Thus, the HLA-B27 allele contributed to the radiographic features in 44% of patients with rheumatoid arthritis and associated HLA-B27. Thus, the wide range of findings in our population indicates that the radiographic attributes are not specific enough to constitute a unique subpopulation of patients with rheumatoid arthritis. (orig.)

Rundback, J.H. (Dept. of Radiology, Beth Israel Medical Center, New York, NY (United States)); Rosenberg, Z.S. (Dept. of Radiology, Hospital for Joint Diseases, Orthopaedic Inst., New York, NY (United States)); Solomon, G. (Dept. of Rheumatology, Hospital for Joint Diseases, Orthopaedic Institute, New York, NY (United States))

1993-05-01

34

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

Science.gov (United States)

HLA-B27 (B27) is strongly associated with the spondyloarthritides. B27 is expressed at the cell surface of antigen presenting cells (APC) both as canonical ?2m-associated and non-canonical ?2m-free heavy chain (FHC) forms which include B27 dimers (termed B272). B27 FHC forms arise in an endosomal compartment from recycling ?2m-associated B27. Formation of cell surface FHC dimers is critically dependent on an unpaired reactive cysteine 67 in the ?1 helix of the class I heavy chain. HLA-B27 also form redox-inducible ?2m-associated dimers on exosomes and apoptosing cells. By contrast with cell surface expressed cysteine 67-dependent heavy chain dimers these dimers are dependent on a cytoplasmic cysteine 325 for their formation. HLA-B27 binds to immunoregulatory receptors including members of the Killer cell Immunoglobulin-like (KIR) and Leukocyte Immunoglobulin-like receptor family. B27 FHC bind to different but overlapping sets of these immunoreceptors compared to classical ?2m-associated HLA-B27. B27 FHC bind more strongly to KIR3DL2 and LILRB2 immune receptor than other ?2m-associated HLA-class I ligands. Genetic studies have implicated genes which control production of the important proinflammatory cytokine IL-17 in the pathogenesis of spondyloarthritis. Cell surface HLA-B27 FHC binding to these immune receptors or acting through other mechanisms could impact on the pathogenesis of spondyloarthritis by promoting immune cell production of IL-17. Here we review the literature on these non-canonical forms of HLA-B27 and the immune receptors they bind to and discuss the possible relevance of these interactions to the pathogenesis of spondyloarthropathy. PMID:23910730

Shaw, Jacqueline; Hatano, Hiroko; Kollnberger, Simon

2013-08-01

35

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

UK PubMed Central (United Kingdom)

HLA-B27 (B27) is strongly associated with the spondyloarthritides. B27 is expressed at the cell surface of antigen presenting cells (APC) both as canonical ?2m-associated and non-canonical ?2m-free heavy chain (FHC) forms which include B27 dimers (termed B272). B27 FHC forms arise in an endosomal compartment from recycling ?2m-associated B27. Formation of cell surface FHC dimers is critically dependent on an unpaired reactive cysteine 67 in the ?1 helix of the class I heavy chain. HLA-B27 also form redox-inducible ?2m-associated dimers on exosomes and apoptosing cells. By contrast with cell surface expressed cysteine 67-dependent heavy chain dimers these dimers are dependent on a cytoplasmic cysteine 325 for their formation. HLA-B27 binds to immunoregulatory receptors including members of the Killer cell Immunoglobulin-like (KIR) and Leukocyte Immunoglobulin-like receptor family. B27 FHC bind to different but overlapping sets of these immunoreceptors compared to classical ?2m-associated HLA-B27. B27 FHC bind more strongly to KIR3DL2 and LILRB2 immune receptor than other ?2m-associated HLA-class I ligands. Genetic studies have implicated genes which control production of the important proinflammatory cytokine IL-17 in the pathogenesis of spondyloarthritis. Cell surface HLA-B27 FHC binding to these immune receptors or acting through other mechanisms could impact on the pathogenesis of spondyloarthritis by promoting immune cell production of IL-17. Here we review the literature on these non-canonical forms of HLA-B27 and the immune receptors they bind to and discuss the possible relevance of these interactions to the pathogenesis of spondyloarthropathy.

Shaw J; Hatano H; Kollnberger S

2014-01-01

36

Detection of HLA-B*27 gene using a spectral plasmon resonance imaging system.  

UK PubMed Central (United Kingdom)

Detection of HLA-B*27 gene is clinically important due to its strong association with diseases, such as ankylosing spondylitis. Nucleic acid-based biosensors represent a promising clinical tool for gene diagnosis. Surface plasmon resonance (SPR) can be used to monitor DNA-DNA hybridization in real-time and without any prior labeling step. Here, a self-built HLA-B*27 positive PCR products spectral SPR imaging system was used for multichannel direct-detection of a specific sequence of the HLA-B*27 gene. Thiol-labeled single-stranded oligonucleotide probes, which were proved to be superior to amine-labeled probes in immobilization, were immobilized onto the surfaces of the gold spots on the sensor chip to target the specific sequence in the HLA-B*27 gene in blood. SPR measurements were performed with different concentrations of synthetic target DNA sequence. The calibration curve of synthetic target sequence showed a good linear relationship between the concentration of the synthetic target sequence and the shift of the SPR wavelength from 10 nM to 500 nM with a detection limit of 5 nM. The HLA-B*27 gene was isolated from human whole blood and amplified using polymerase chain reaction (PCR). PCR products were measured using the SPR imaging system. HLA-B*27 positive PCR products showed significant SPR wavelength shift, while synthetic non-complementary sequence and negative PCR products showed no significant SPR wavelength shift. The method is high-specific, high-throughput and label-free.

Yang Y; Yuan L; Fang X; Liang X; Yang F

2013-08-01

37

HLA-B27 (antigen) in retroperitoneal fibrosis in a family  

Directory of Open Access Journals (Sweden)

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

Mohammad Yazdani; Afshin Shadmehr

2007-01-01

38

Association between polymorphism of the vitamin D metabolism gene CYP27B1 and HLA-B27-associated uveitis. Is a state of relative immunodeficiency pathogenic in HLA B27-positive uveitis?  

UK PubMed Central (United Kingdom)

OBJECTIVE: Polymorphisms of the vitamin D metabolism gene CYP27B1 showed associations with multiple autoimmune diseases. The aim of this study was to investigate a possible association between the rs703842 A>G polymorphism of the CYP27B1 gene and HLA-B27-associated uveitis. DESIGN: One hundred fifty-nine patients with HLA-B27-associated uveitis, 138 HLA-B27-negative controls and 100 HLA-B27-positive controls were recruited for this retrospective case-control study. Main outcome parameters were genotype distribution and allelic frequencies determined by polymerase chain reaction. RESULTS: Carriers of the rs703842G allele were found significantly more often in patients with HLA-B27-associated uveitis than in HLA-B27-positive controls (p = 0.03). Between patients and HLA-B27-negative controls no significant difference in the genotype distribution of the rs703842 A>G polymorphism was found (p = 0.97). CONCLUSIONS: Our data suggest that the rs703842 A>G polymorphism may play a role in HLA-B27-associated uveitis.

Steinwender G; Lindner E; Weger M; Plainer S; Renner W; Ardjomand N; El-Shabrawi Y

2013-01-01

39

The expression of HLA-B27 locus-associated antigens in ankylosing spondylitis  

UK PubMed Central (United Kingdom)

To study the expression of HLA-B27 locus-associated antigens in patients with ankylosing spondylitis(AS). The HLA-B27 associated antigens were exiamined in 418 patients with spondyloarthropathies(SpAs) and 30 controls individuals by complement dependent microcytotoxity assay. There were 73 patients with AS in the SpAs. The percentage of positive Bw6,B27(47),B27 and B7/B27/B73/ B81 were 31%,28%,25% and 22%,respectively. In both the HLA-B27-postive group and HLA-B27-negative group, there were significantly difference among B27 alleles antigens of B27(47),B27 and B7.There was negatively correlation in both antigens B13 and Bw6.Conversely,there was positively correlation in both match antigens of Bw4 and B27(47),B7 and B27. There were strongly linkage disequilibrium in five match antigens of B13 and Bw6,B13 and B60/61/47/48/81(13),B7 and B27,Bw4 and B27(47),Cw1 and B42/B45. B27 antigens are not the only a factor to determine susceptibility to AS, other alleles-associated antigens may be either enhancing,such as Cw1 and Cw2 or suppressive,on the development of AS.

Jiang Yuzhang; Chen Caiyun; Liu Liang

40

HLA-B27 and antigen presentation: At the crossroads between immune defense and autoimmunity.  

UK PubMed Central (United Kingdom)

The HLA-B27 is historically studied as a susceptibility factor in spondyloarthropathies and, primarily, in ankylosing spondylitis (AS). Over the recent years however, it has been rediscovered as protective factor against some severe viral infections. This is due to the high capacity of virus-specific, HLA-B27-restricted CD8+ T cells for both intrinsic (i.e. polyfunctionality, high avidity, low sensitivity to Treg cell-mediated suppression) and extrinsic (i.e. rapid and efficient antigen processing and presentation) factors. It is tempting to speculate that these two aspects are not independent and that the association of B27 molecules to autoimmunity is the downside of this superior functional efficacy which, in given genetic backgrounds and environmental conditions, can support a chronic inflammation leading to spondyloarthropathies. Still, the pathogenic role of HLA-B27 molecules in AS is elusive. Here, we focus on the biology of HLA-B27 from the genetics to the biochemistry and to the structural/dynamical properties of B27:peptide complexes as obtained from atomistic molecular dynamics simulation. Overall, the results point at the antigen presentation as the key event in the disease pathogenesis. In particular, an extensive comparison of HLA-B*2705 and B*2709 molecules, that differ in a single amino acid (Asp116 to His116) and are differentially associated with AS, indicates that position 116 is crucial for shaping the entire peptide-presenting groove.

Sorrentino R; Böckmann RA; Fiorillo MT

2014-01-01

 
 
 
 
41

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

International Nuclear Information System (INIS)

[en] 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

42

Functional interaction of the ankylosing spondylitis-associated endoplasmic reticulum aminopeptidase 1 polymorphism and HLA-B27 in vivo.  

UK PubMed Central (United Kingdom)

The association of ERAP1 with ankylosing spondylitis (AS)1 among HLA-B27-positive individuals suggests that ERAP1 polymorphism may affect pathogenesis by altering peptide-dependent features of the HLA-B27 molecule. Comparisons of HLA-B*27:04-bound peptidomes from cells expressing different natural variants of ERAP1 revealed significant differences in the size, length, and amount of many ligands, as well as in HLA-B27 stability. Peptide analyses suggested that the mechanism of ERAP1/HLA-B27 interaction is a variant-dependent alteration in the balance between epitope generation and destruction determined by the susceptibility of N-terminal flanking and P1 residues to trimming. ERAP1 polymorphism associated with AS susceptibility ensured efficient peptide trimming and high HLA-B27 stability. Protective polymorphism resulted in diminished ERAP1 activity, less efficient trimming, suboptimal HLA-B27 peptidomes, and decreased molecular stability. This study demonstrates that natural ERAP1 polymorphism affects HLA-B27 antigen presentation and stability in vivo and proposes a mechanism for the interaction between these molecules in AS.

García-Medel N; Sanz-Bravo A; Van Nguyen D; Galocha B; Gómez-Molina P; Martín-Esteban A; Alvarez-Navarro C; de Castro JA

2012-11-01

43

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

Directory of Open Access Journals (Sweden)

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.

Samuel A. Sakyi; Margaret T. Agyei-Frempong; Robert E. Quansah

2012-01-01

44

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

Directory of Open Access Journals (Sweden)

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

Wexler Alexandra; Sand Trond; Elsås Tor B

2012-01-01

45

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

UK PubMed Central (United Kingdom)

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

Wexler A; Sand T; Elsås TB

2012-01-01

46

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

DEFF Research Database (Denmark)

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

Berntson, Lillemor; Damgård, Michael

2008-01-01

47

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

48

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

49

Phosphorylation of STAT-1 serine 727 is prolonged in HLA-B27-expressing human monocytic cells.  

Science.gov (United States)

A tissue antigen, HLA-B27, is strongly associated with a group of rheumatic diseases called spondyloarthritides. Despite the intensive research, the exact role of HLA-B27 in the pathogenesis of these diseases is still unclear. Here we studied whether HLA-B27 modulates the phosphorylation of signal transducer and activator of transcription 1 (STAT-1) serine 727 residue and the localization of STAT-1 in Salmonella-infected human monocytic cells. In addition, we studied the role of signaling molecule double-stranded RNA activated protein kinase (PKR) in these modulatory effects. U937 human monocytic cell transfectants stably expressing wild type HLA-B27 or mutated HLA-B27 heavy chains with amino acid substitutions in the B pocket were prepared. The PMA-differentiated cells were infected with S. enteritidis. Western blotting was used to detect the phosphorylation of STAT-1, and to visualize the localization of STAT-1 in the cells confocal microscopy was used. Specific inhibitors were employed to study the role of PKR in STAT-1 phosphorylation. We discovered that the phosphorylation of STAT-1 serine 727 is prolonged in cells expressing misfolding forms of HLA-B27 after S. enteritidis infection, whereas in mock cells and in cells expressing mutated, non-misfolding HLA-B27 the phosphorylation of serine 727 is transient. Interestingly, STAT-1 serine 727 phosphorylation is partly dependent on PKR. In addition, more STAT-1 is localized in the nucleus of HLA-B27-expressing cells, even before an external trigger, when compared to mock cells. In conclusion, our results show that the phosphorylation of STAT-1 serine 727 residue is prolonged in HLA-B27-expressing monocyte-macrophage U937 cells after bacterial infection. This is of interest since the phosphorylation of serine 727 on STAT-1 is suggested to contribute to macrophage activation and promote inflammatory responses. Therefore, our results provide a mechanism which explains how the expression of an HLA-B27 molecule can impact the course of Salmonella infection and reactive arthritis. PMID:23349666

Ruuska, Marja; Sahlberg, Anna S; Granfors, Kaisa; Penttinen, Markus A

2013-01-21

50

Phosphorylation of STAT-1 serine 727 is prolonged in HLA-B27-expressing human monocytic cells.  

UK PubMed Central (United Kingdom)

A tissue antigen, HLA-B27, is strongly associated with a group of rheumatic diseases called spondyloarthritides. Despite the intensive research, the exact role of HLA-B27 in the pathogenesis of these diseases is still unclear. Here we studied whether HLA-B27 modulates the phosphorylation of signal transducer and activator of transcription 1 (STAT-1) serine 727 residue and the localization of STAT-1 in Salmonella-infected human monocytic cells. In addition, we studied the role of signaling molecule double-stranded RNA activated protein kinase (PKR) in these modulatory effects. U937 human monocytic cell transfectants stably expressing wild type HLA-B27 or mutated HLA-B27 heavy chains with amino acid substitutions in the B pocket were prepared. The PMA-differentiated cells were infected with S. enteritidis. Western blotting was used to detect the phosphorylation of STAT-1, and to visualize the localization of STAT-1 in the cells confocal microscopy was used. Specific inhibitors were employed to study the role of PKR in STAT-1 phosphorylation. We discovered that the phosphorylation of STAT-1 serine 727 is prolonged in cells expressing misfolding forms of HLA-B27 after S. enteritidis infection, whereas in mock cells and in cells expressing mutated, non-misfolding HLA-B27 the phosphorylation of serine 727 is transient. Interestingly, STAT-1 serine 727 phosphorylation is partly dependent on PKR. In addition, more STAT-1 is localized in the nucleus of HLA-B27-expressing cells, even before an external trigger, when compared to mock cells. In conclusion, our results show that the phosphorylation of STAT-1 serine 727 residue is prolonged in HLA-B27-expressing monocyte-macrophage U937 cells after bacterial infection. This is of interest since the phosphorylation of serine 727 on STAT-1 is suggested to contribute to macrophage activation and promote inflammatory responses. Therefore, our results provide a mechanism which explains how the expression of an HLA-B27 molecule can impact the course of Salmonella infection and reactive arthritis.

Ruuska M; Sahlberg AS; Granfors K; Penttinen MA

2013-01-01

51

Implications of structural and thermodynamic studies of HLA-B27 subtypes exhibiting differential association with ankylosing spondylitis.  

UK PubMed Central (United Kingdom)

Structural and thermodynamic properties of HLA-B27 molecules provide the basis for their function within the immune system and are probably also central for the understanding of the pathology of HLA-B27-associated diseases such as ankolysing spondylitis (AS). Several HLA-B27 alleles are AS-associated, whereas some are not, although the protein encoded by the former may differ in only a single amino acid exchange from those specified by the latter. This indicates that subtype-specific polymorphic residues play a key role in determining whether an HLA-B27 subtype is AS-associated or not and open the possibility to correlate structural, thermodynamic and functional characteristics ofa given subtype with the disease association. Our studies involved X-ray crystallography and various other biophysical techniques to examine how several different peptides are accommodated within the binding groove of the molecules. The HLA-B*2705 and HLA-B*2709 subtypes, whose products differ in only a single amino acid residue of their heavy chains from each other, were primarily chosen for these analyses, but our studies have recently also been extended to the closely related subtypes HLA-B*2703, HLA-B*2704 and HLA-B*2706. The analyses reveal that structural and thermodynamic differences between HLA-B27 complexes may exist, depending on the peptide that is displayed. Furthermore, aviralpeptide and two self-peptides were found that exhibit HLA-B27 subtype-dependent molecular mimicry, thereby providing a molecular basis to account for the subtype-dependent presence of autoreactive T-cells. Although these results do not exclude other theories for the pathogenesis of AS, they support the arthritogenic peptide hypothesis which envisages molecular mimicry between HLA-B27-presented foreign and self-peptides to explain the cross-reactivity of autoreactive T-cells that are found in HLA-B*2705-positive individuals, in particular when they suffer from AS.

Ziegler A; Loll B; Misselwitz R; Uchanska-Ziegler B

2009-01-01

52

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

DEFF Research Database (Denmark)

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

Schiellerup, P.; Krogfelt, K.A.

2008-01-01

53

Altered regulation of ELAVL1/HuR in HLA-B27-expressing U937 monocytic cells.  

UK PubMed Central (United Kingdom)

OBJECTIVE: To investigate the role of HLA-B27 expression in the regulation of RNA binding protein (RBP) Embryonic Lethal Abnormal Vision (ELAV) L1/Human antigen R (HuR) expression in Salmonella-infected or LPS-stimulated human monocytic cells, since HuR is a critical regulator of the post-transcriptional fate of many genes (e.g. TNF?) important in inflammatory response. METHODS: U937 monocytic cells were stably transfected with pSV2neo resistant vector (mock), wild type HLA-B27, or mutated HLA-B27 with amino acid substitutions in the B pocket. Cells were differentiated, infected with Salmonella enteritidis or stimulated with lipopolysaccharide. The expression levels of HuR protein and cleavage products (CP1 and CP2) were detected by Western blotting and flow cytometry. Specific inhibitors were used to study the role of PKR and p38 in HuR expression and generation of CPs. TNF? and IL-10 secretion after p38 and PKR inhibition were measured by ELISA. RESULTS: Full length HuR is overexpressed and HuR cleavage is disturbed in U937 monocytic cells expressing HLA-B27 heavy chains (HC). Increased full length HuR expression, disturbed cleavage and reduced dependence on PKR after infection correlate with the expression of glutamic acid 45 in the B pocket that is linked to the misfolding of HLA-B27. CONCLUSION: Results show that the expression of HLA-B27 HCs modulates the intracellular environment of U937 monocyte/macrophages by altering HuR regulation. This phenomenon is at least partly dependent on the misfolding feature of the B27 molecule. Since HuR is an important regulator of multiple genes involved in inflammatory response observations offer an explanation how HLA-B27 may modulate inflammatory response.

Sahlberg AS; Ruuska M; Granfors K; Penttinen MA

2013-01-01

54

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

Directory of Open Access Journals (Sweden)

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

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

2007-01-01

55

HLA-B?27 subtype specificity determines targeting and viral evolution of a hepatitis C virus-specific CD8+ T-cell epitope.  

UK PubMed Central (United Kingdom)

BACKGROUND & AIMS: HLA-B?27 is associated with spontaneous HCV genotype 1 clearance. HLA-B?27-restricted CD8+ T-cells target three NS5B epitopes. Two of these epitopes are dominantly targeted in the majority of HLA-B?27+ patients. In chronic infection, viral escape occurs consistently in these two epitopes. The third epitope (NS5B2820) was dominantly targeted in an acutely infected patient. This was in contrast, however, to the lack of recognition and viral escape in the large majority of HLA-B?27+ patients. Here, we set out to determine the host factors contributing to selective targeting of this epitope. METHODS: Four-digit HLA class I typing and viral sequence analyses were performed in 78 HLA-B?27+ patients with chronic HCV genotype 1 infection. CD8+ T-cell analyses were performed in a subset of patients. In addition, HLA/peptide affinity was compared for HLA-B?27:02 and 05. RESULTS: The NS5B2820 epitope is only restricted by the HLA-B?27 subtype HLA-B?27:02 (that is frequent in Mediterranean populations), but not by the prototype HLA-B?27 subtype B?27:05. Indeed, the epitope is very dominant in HLA-B?27:02+ patients and is associated with viral escape mutations at the anchor position for HLA-binding in 12 out of 13 HLA-B?27:02+ chronically infected patients. CONCLUSIONS: The NS5B2820 epitope is immunodominant in the context of HLA-B?27:02, but is not restricted by other HLA-B?27 subtypes. This finding suggests an important role of HLA subtypes in the restriction of HCV-specific CD8+ responses. With minor HLA subtypes covering up to 39% of specific populations, these findings may have important implications for the selection of epitopes for global vaccines. (250/250).

Nitschke K; Barriga A; Schmidt J; Timm J; Viazov S; Kuntzen T; Kim AY; Lauer GM; Allen TM; Gaudieri S; Rauch A; Lange CM; Sarrazin C; Eiermann T; Sidney J; Sette A; Thimme R; López D; Neumann-Haefelin C

2013-08-01

56

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

Science.gov (United States)

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

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

1998-10-01

57

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

UK PubMed Central (United Kingdom)

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

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

1998-10-01

58

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

DEFF Research Database (Denmark)

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

Boucherma, Rachid; Kridane-Miledi, Hédia

2013-01-01

59

HLA-B27 predicts a more chronic disease course in an 8-year followup cohort of patients with juvenile idiopathic arthritis.  

UK PubMed Central (United Kingdom)

OBJECTIVE: We investigated associations of HLA-B27 with clinical manifestations and longterm outcome in a near population-based setting among patients with juvenile idiopathic arthritis (JIA). METHODS: We studied clinical and serological data from 410 patients with HLA-B27 results among 440 prospectively collected patients with JIA with 8-year followup data in a Nordic database. The study was structured to be as close to a population-based study as possible. RESULTS: HLA-B27 was analyzed in 93% of patients, and was positive in 21% of the cohort, in 18.4% of the girls and in 25.9% of the boys. Boys who were HLA-B27-positive had significantly higher age at onset compared to HLA-B27-negative boys and compared to both HLA-B27-negative and positive girls. This difference in onset age in relation to HLA-B27 was not found in girls. HLA-B27 was associated with clinical signs of sacroiliitis, enthesitis, and tenosynovitis in boys, but not in girls. After 8 years of disease, 46 children (11.2%) were classified as having enthesitis-related arthritis (ERA). Boys with ERA had clinical signs of sacroiliitis more often than girls with ERA. HLA-B27-positive children, as well as children with clinical signs of sacroiliitis, enthesitis, and hip arthritis, had higher odds of not being in remission off medication after 8 years of disease. CONCLUSION: In this near population-based Nordic JIA cohort we found significant differences between HLA-B27-positive boys and girls in age at disease onset, clinical signs of sacroiliitis, and ERA classification. HLA-B27 was negatively associated with longterm remission status, possibly because of its association with clinical disease characteristics, such as sacroiliitis, rather than being a general marker of persistent disease.

Berntson L; Nordal E; Aalto K; Peltoniemi S; Herlin T; Zak M; Nielsen S; Rygg M

2013-05-01

60

Novel HLA-B27-restricted epitopes from Chlamydia trachomatis generated upon endogenous processing of bacterial proteins suggest a role of molecular mimicry in reactive arthritis.  

UK PubMed Central (United Kingdom)

Reactive arthritis (ReA) is an HLA-B27-associated spondyloarthropathy that is triggered by diverse bacteria, including Chlamydia trachomatis, a frequent intracellular parasite. HLA-B27-restricted T-cell responses are elicited against this bacterium in ReA patients, but their pathogenetic significance, autoimmune potential, and relevant epitopes are unknown. High resolution and sensitivity mass spectrometry was used to identify HLA-B27 ligands endogenously processed and presented by HLA-B27 from three chlamydial proteins for which T-cell epitopes were predicted. Fusion protein constructs of ClpC, (Na+)-translocating NADH-quinone reductase subunit A, and DNA primase were expressed in HLA-B27+ cells and their HLA-B27-bound peptidomes were searched for endogenous bacterial ligands. A non-predicted peptide, distinct from the predicted T-cell epitope, was identified from ClpC. A peptide recognized by T cells in vitro, NQRA(330-338), was detected from the reductase subunit. This is the second HLA-B27-restricted T-cell epitope from C. trachomatis with relevance in ReA demonstrated to be processed and presented in live cells. A novel peptide from the DNA primase, DNAP(211-223), was also found. This was a larger variant of a known epitope and was highly homologous to a self-derived natural ligand of HLA-B27. All three bacterial peptides showed high homology with human sequences containing the binding motif of HLA-B27. Molecular dynamics simulations further showed a striking conformational similarity between DNAP(211-223) and its homologous and much more flexible human-derived HLA-B27 ligand. The results suggest that molecular mimicry between HLA-B27-restricted bacterial and self-derived epitopes is frequent and may play a role in ReA.

Alvarez-Navarro C; Cragnolini JJ; Dos Santos HG; Barnea E; Admon A; Morreale A; López de Castro JA

2013-07-01

 
 
 
 
61

Novel HLA-B27-restricted Epitopes from Chlamydia trachomatis Generated upon Endogenous Processing of Bacterial Proteins Suggest a Role of Molecular Mimicry in Reactive Arthritis.  

Science.gov (United States)

Reactive arthritis (ReA) is an HLA-B27-associated spondyloarthropathy that is triggered by diverse bacteria, including Chlamydia trachomatis, a frequent intracellular parasite. HLA-B27-restricted T-cell responses are elicited against this bacterium in ReA patients, but their pathogenetic significance, autoimmune potential, and relevant epitopes are unknown. High resolution and sensitivity mass spectrometry was used to identify HLA-B27 ligands endogenously processed and presented by HLA-B27 from three chlamydial proteins for which T-cell epitopes were predicted. Fusion protein constructs of ClpC, Na(+)-translocating NADH-quinone reductase subunit A, and DNA primase were expressed in HLA-B27(+) cells, and their HLA-B27-bound peptidomes were searched for endogenous bacterial ligands. A non-predicted peptide, distinct from the predicted T-cell epitope, was identified from ClpC. A peptide recognized by T-cells in vitro, NQRA(330-338), was detected from the reductase subunit. This is the second HLA-B27-restricted T-cell epitope from C. trachomatis with relevance in ReA demonstrated to be processed and presented in live cells. A novel peptide from the DNA primase, DNAP(211-223), was also found. This was a larger variant of a known epitope and was highly homologous to a self-derived natural ligand of HLA-B27. All three bacterial peptides showed high homology with human sequences containing the binding motif of HLA-B27. Molecular dynamics simulations further showed a striking conformational similarity between DNAP(211-223) and its homologous and much more flexible human-derived HLA-B27 ligand. The results suggest that molecular mimicry between HLA-B27-restricted bacterial and self-derived epitopes is frequent and may play a role in ReA. PMID:23867464

Alvarez-Navarro, Carlos; Cragnolini, Juan J; Dos Santos, Helena G; Barnea, Eilon; Admon, Arie; Morreale, Antonio; López de Castro, José A

2013-07-18

62

Novel HLA-B27-restricted epitopes from Chlamydia trachomatis generated upon endogenous processing of bacterial proteins suggest a role of molecular mimicry in reactive arthritis.  

UK PubMed Central (United Kingdom)

Reactive arthritis (ReA) is an HLA-B27-associated spondyloarthropathy that is triggered by diverse bacteria, including Chlamydia trachomatis, a frequent intracellular parasite. HLA-B27-restricted T-cell responses are elicited against this bacterium in ReA patients, but their pathogenetic significance, autoimmune potential, and relevant epitopes are unknown. High resolution and sensitivity mass spectrometry was used to identify HLA-B27 ligands endogenously processed and presented by HLA-B27 from three chlamydial proteins for which T-cell epitopes were predicted. Fusion protein constructs of ClpC, Na(+)-translocating NADH-quinone reductase subunit A, and DNA primase were expressed in HLA-B27(+) cells, and their HLA-B27-bound peptidomes were searched for endogenous bacterial ligands. A non-predicted peptide, distinct from the predicted T-cell epitope, was identified from ClpC. A peptide recognized by T-cells in vitro, NQRA(330-338), was detected from the reductase subunit. This is the second HLA-B27-restricted T-cell epitope from C. trachomatis with relevance in ReA demonstrated to be processed and presented in live cells. A novel peptide from the DNA primase, DNAP(211-223), was also found. This was a larger variant of a known epitope and was highly homologous to a self-derived natural ligand of HLA-B27. All three bacterial peptides showed high homology with human sequences containing the binding motif of HLA-B27. Molecular dynamics simulations further showed a striking conformational similarity between DNAP(211-223) and its homologous and much more flexible human-derived HLA-B27 ligand. The results suggest that molecular mimicry between HLA-B27-restricted bacterial and self-derived epitopes is frequent and may play a role in ReA.

Alvarez-Navarro C; Cragnolini JJ; Dos Santos HG; Barnea E; Admon A; Morreale A; López de Castro JA

2013-09-01

63

Aetiology and pathogenesis of reactive arthritis: role of non-antigen-presenting effects of HLA-B27  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Spondyloarthropathies are inflammatory diseases closely associated with human leukocyte antigen (HLA)-B27 by unknown mechanisms. One of these diseases is reactive arthritis (ReA), which is typically triggered by Gram-negative bacteria, which have lipopolysaccharide as an integral component of their ...

Vähämiko, Sanna; Penttinen, Markus A; Granfors, Kaisa

64

HLA-B*27:05-specific cytotoxic T lymphocyte epitopes in Indian HIV type 1C.  

UK PubMed Central (United Kingdom)

HLA-B*27:05 is one of the widely reported alleles associated with resistance to HIV, while HLA-A24, HLA-B7, HLA-B*07:02, HLA-B*35:01, HLA-B*53:01, and HLA-B40 are reported to be associated with susceptibility to HIV. Using a bioinformatics approach we attempted to predict potential HLA-B*27:05-specific HIV-1C epitopes that do not bind to susceptibility-associated HLA alleles based on our hypothesis that such epitopes have a greater probability of eliciting a protective immune response in the host. A consensus sequence was built for all proteins of Indian clade C virus. Epitopes specific to HLA-B*27:05 were predicted from the consensus sequence using two different bioinformatics methods to enhance the accuracy of the prediction. Epitopes that were also predicted to bind to any of the susceptibility-associated HLA alleles were excluded from the list. The short-listed epitopes were modeled using MODPROPEP to refine the prediction. Fourteen peptides were identified as epitopes by both sequence-based methods and were found to interact strongly with HLA-B*27:05 by molecular modeling studies. Five of the 14 epitopes were previously reported as immunogenic by other researchers, while the remaining nine are novel. The 14 epitopes have been repeatedly identified by three different methods indicating their potential as useful candidates for an effective HIV vaccine.

Sundaramurthi JC; Ramanathan VD; Hanna LE

2013-01-01

65

Implications of structural and thermodynamic studies of HLA-B27 subtypes exhibiting differential association with ankylosing spondylitis.  

Science.gov (United States)

Structural and thermodynamic properties of HLA-B27 molecules provide the basis for their function within the immune system and are probably also central for the understanding of the pathology of HLA-B27-associated diseases such as ankolysing spondylitis (AS). Several HLA-B27 alleles are AS-associated, whereas some are not, although the protein encoded by the former may differ in only a single amino acid exchange from those specified by the latter. This indicates that subtype-specific polymorphic residues play a key role in determining whether an HLA-B27 subtype is AS-associated or not and open the possibility to correlate structural, thermodynamic and functional characteristics ofa given subtype with the disease association. Our studies involved X-ray crystallography and various other biophysical techniques to examine how several different peptides are accommodated within the binding groove of the molecules. The HLA-B*2705 and HLA-B*2709 subtypes, whose products differ in only a single amino acid residue of their heavy chains from each other, were primarily chosen for these analyses, but our studies have recently also been extended to the closely related subtypes HLA-B*2703, HLA-B*2704 and HLA-B*2706. The analyses reveal that structural and thermodynamic differences between HLA-B27 complexes may exist, depending on the peptide that is displayed. Furthermore, aviralpeptide and two self-peptides were found that exhibit HLA-B27 subtype-dependent molecular mimicry, thereby providing a molecular basis to account for the subtype-dependent presence of autoreactive T-cells. Although these results do not exclude other theories for the pathogenesis of AS, they support the arthritogenic peptide hypothesis which envisages molecular mimicry between HLA-B27-presented foreign and self-peptides to explain the cross-reactivity of autoreactive T-cells that are found in HLA-B*2705-positive individuals, in particular when they suffer from AS. PMID:19731629

Ziegler, Andreas; Loll, Bernhard; Misselwitz, Rolf; Uchanska-Ziegler, Barbara

2009-01-01

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

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Chlamydia trachomatis triggers reactive arthritis, a spondyloarthropathy linked to the human major histocompatibility complex molecule HLA-B27, through an unknown mechanism that might involve molecular mimicry between chlamydial and self-derived HLA-B27 ligands. Chlamydia-specific CD8+ T-cells are f...

Cragnolini, Juan J.; García-Medel, Noel; López de Castro, José A.

67

Association of ankylosing spondylitis with HLA-B27 and ERAP1: pathogenic role of antigenic peptide.  

UK PubMed Central (United Kingdom)

Ankylosing spondylitis (AS) is a form of seronegative inflammatory arthritis whose strong genetic association with the human leucocyte antigen (HLA)-B27 has been known for almost 4 decades. However, its mechanism remains poorly understood. Recently, with the development of genetics, further more genes have been robustly associated with the disease. Genome-wide association studies identified the association between AS and ERAP1 (endoplasmic reticulum associated aminopeptidase 1). And ERAP1 has shown the potential in trimming antigenic peptides to optimal length for binding to HLA-B27 in the ER (endoplasmic reticulum). However, the length of the peptides are strictly restricted in the process of peptide transporting, processing and presentation. A hypothesis is proposed that the abnormal mechanism of AS may related to the trimming of N-terminal sequences from antigenic precursors in the ER and the length of the antigenic peptides that are presented to the T-cell receptors.

Chen B; Li D; Xu W

2013-01-01

68

[Relation of spondylarthropathies and HLA-B27 antigen in patients from the state of Zulia, Venezuela  

UK PubMed Central (United Kingdom)

HLA-B27 and associate antigens incidence were studied in 620 cases of seronegative spondiloarthropathies (SNS) and 262 controls of a Venezuelan mestizo population from Zulla state between 1985 and 1995. The incidence of HLA-B27 was 20.96% of all cases of SNS. It was increased in patients with ankilosing spondylitis (AS) 33.33% and Relter's syndrome (RS) 30%, but not in uveitis (Uv) 20% an psoriatic arthropathy (PsA) 0%. The incidence in the control group was 4.2%. This results are lower than the previous description in Venezuelan mestizo and caucasic population but are close to the incidence described in population of West Africa with important contribution to admixture of the occidental part of Venezuela.

Rivera S; Hassanhi M; Márquez G; Fuenmayor A; Monzón J; Avila J

1996-12-01

69

[Chlamydia-induced reactive arthritis--HLA-B 27 negative two patients  

UK PubMed Central (United Kingdom)

Two cases with HLA-B 27 negative, Chlamydia-induced reactive arthritis (ReA) were described. Case 1: A 30 y.o. male developed balanitis, urethritis, arthritis of both knees, elbows, shoulders and hip joints on May in 1997. Laboratory findings revealed CRP 2.7 mg/dl (normal range < 0.3), ESR 33 mm/h and negative rheumatoid factor (RF) test. Anti-Chlamydia trachomatis antibodies, IgG 2.22, IgA 3.33 were positive. HLA-typing revealed A 2, A 24 (9), B 39 (16), B 52 (5). He was diagnosed as ReA and arthritis subsided with treatment of minocycline and nonsteroidal antiinflammatory drugs (NSAIDs). Case 2: A 40 y.o. Iranian American male developed balanitis, urethritis, lumbago, arthritis of both elbows, knees and foot joints, iridocyclitis on August in 1995. Chlamydia trachomatis was detected in the urethral swab culture. He was diagnosed as ReA and treated with minocycline and NSAIDs. He was referred to our hospital on June in 1996. Arthritis at both knees and feet was detected. Laboratory findings revealed CRP 0.8 mg/dl, negative RF test was revealed. Antibodies to Chlamydia were positive (IgG 1.49, IgA 1.53) positive. HLA typing revealed A 1, A 2, B 37, B 55 (22). He was again treated with minocycline and NSAIDs and ReA ameliorated. Since HLA-B 22, B 37 and B 39 have been reported to cross-react or to have homology with B 27, B 22, B 37 and B 39 are likely to related to inducing ReA.

Kobayashi S; Suzuki S; Ueda A; Ushiyama C; Tamura N; Inoue H; Tsuda H; Takasaki Y; Hashimoto H

1999-02-01

70

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.

1987-01-01

71

HLA-B27 modulates intracellular growth of Salmonella pathogenicity island 2 mutants and production of cytokines in infected monocytic U937 cells.  

UK PubMed Central (United Kingdom)

BACKGROUND: Salmonella enterica serovar Enteritidis PT4 KS8822/88 replicates rapidly in HLA-B27-transfected human monocytic U937 cells. In this process, Salmonella pathogenicity island 2 (SPI-2) genes play a crucial role. Our previous study indicated that 118 Salmonella genes, including 8 SPI-2 genes were affected by HLA-B27 antigen during Salmonella infection of U937 cells. METHODS/PRINCIPAL FINDINGS: To further investigate Salmonella replication in HLA-B27-positive U937 monocytic cells, two SPI-2 genes, ssaS and sscA up-regulated most during Salmonella infection of HLA-B27-transfected U937 cells, were mutated by using one-step gene disruption method. Intracellular survival and replication of the mutants in the U937 cells was compared to that of the wild type strain. Surprisingly, the two mutated strains replicated significantly more than the wild type bacteria in HLA-B27-transfected cells. Secretion of tumor necrosis factor alpha (TNF-?) and interleukin 10 (IL-10) was significantly induced during the infection of HLA-B27-transfected U937 cells with the mutants. The results indicated that the certain SPI-2 genes in wild type bacteria suppress Salmonella intracellular growth and production of cytokines in infected HLA-B27-transfected cells. HLA-B27-associated modulation of Salmonella SPI-2 genes and cytokine production may have importance in the persistent infection of the bacteria and the pathogenesis of reactive arthritis. CONCLUSIONS: The study provides evidence that certain virulence factors of pathogens can reduce the intracellular growth in the host cells. We suggest that the limiting intracellular growth might be a strategy for persistence of bacteria in host cells, keeping a balance between pathogenic growth and pathogenesis.

Ge S; He Q; Granfors K

2012-01-01

72

Effect of HLA-B*27 and its Subtypes on Clinical Manifestations and Severity of Ankylosing Spondylitis in Iranian patients.  

UK PubMed Central (United Kingdom)

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.

Fallahi S; Mahmoudi M; Nicknam MH; Gharibdoost F; Farhadi E; Saei A; Nourijelyani K; Ahmadzadeh N; Jamshidi AR

2013-12-01

73

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

2013-08-28

74

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.

Ge Shichao; Danino Vittoria; He Qiushui; Hinton Jay CD; Granfors Kaisa

2010-01-01

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Recognition of B cells epitopes of the Klebsiella pneumoniae GroEL-like protein by HLA-B27 positive subjects.  

UK PubMed Central (United Kingdom)

The presence of antibodies against antigens of K. pneumoniae in HLA-B27 positive patients with ankylosing spondylitis (AS), has been well documented. We have previously reported that sera from HLA-B27 positive subjects react with the K. pneumoniae GroEL-like protein (HSP60Kp) and have higher titers than HLA-B27 negative individuals. We cloned the gene that codes for this protein, determined hydrophilic regions by computer analysis of the predicted amino acid sequence and found that residues 389-397, 360-368 and 282-290, were possible B cell epitopes. To test this prediction, and to determine if the HLA-B27 positive and negative AS patients recognize the same or different epitopes, we truncated the hsp60Kp gene, from the 3; terminal nucleotide, to obtain fragments having or not the predicted epitopes. Four polypeptides of 40, 37, 30 and 18 kDa were obtained and analysed, by ELISA and inhibition of ELISA, for their reactivity with IgG antibodies from three high responders HLA-B27 positive AS patients and three HLA-B27 negative subjects who recognized the rHSP60Kp. Sera from both HLA-B27 positive and negative subjects reacted equally well with rHSP60Kp or with the 40 and 37 kDa peptides, which do not have residues 389-397 and 360-368, respectively, but reactivity was lost with the 30 kDa peptide, which also lacks residues 282-290. Contrary to what we expected, antibodies from HLA-B27 negative and positive individuals recognized the same epitope of the HSP60Kp. Our results indicate that the important epitope for B cells could be the 282-290 region and that the contribution of the two other predicted regions is minimal. We also conclude that the differences in response to the HSP60Kp in HLA-B27 positive AS patients and HLA-B27 negative individuals is not qualitative, but only quantitative.

Cancino-Díaz M; Ayala-Narváez H; Burgos-Vargas R; Selene Reyes-López A; Tovar-Castillo L; Domínguez-López L; Granados Arreola J; Jiménez-Zamudio L; García-Latorre E

2000-04-01

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HLA-B27 homozygosity has no influence on radiographic damage in ankylosing spondylitis: Observation Study of Korean spondyloArthropathy Registry (OSKAR) data.  

UK PubMed Central (United Kingdom)

OBJECTIVES: To evaluate the influence of homozygosity for HLA-B27 on the radiographic damage in ankylosing spondylitis (AS). METHODS: A total of 368 AS patients with positive HLA-B27 status from the Observation Study of Korean spondyloArthropathy Registry (OSKAR) cohort were recruited for this study. HLA-B27 positive patients out of all AS patients were assessed for whether they had homozygosity or heterozygosity for HLA-B27. First, all data were stratified in relation to the carrier state of positive HLA-B27 for cross-sectional survey. Then we compared the radiographic damage score between groups. Second, we evaluated collected clinical and radiographic parameters at two different time points. Then we compared radiographic progression between groups. To use the mSASSS, cervical and lumbar spinal radiographs were examined by two experienced bone and joint radiologists (S. Lee, K.B. Joo). RESULTS: The agreement between the two readers regarding mSASSS was very good: ICC coefficient 0.70 (95% CI 0.60-0.81). The mean age (SD) of the AS patients was 37.0 (9.2) years, and the mean disease duration (SD) was 15.6 (9.1) years. Of these patients, 34.5% (127 patients) had HLA-B27 homozygosity. The mean mSASSS unit (SEM) was not significantly different between groups (homozygosity 28.57±4.12 vs heterozygosity 23.34±3.44, P=0.344) on cross-sectional survey. When it comes to radiographic progression between groups over 5years, there was no significant difference in spite of adjusting for confounding variable (homozygosity 4.98±0.98 vs heterozygosity 4.21±0.82, P=0.562). CONCLUSION: The carrier state of positive HLA-B27 plays no role in determining the radiographic progression in AS.

Kim TJ; Sung IH; Lee S; Joo KB; Choi JH; Park DJ; Park YW; Lee SS; Kim TH

2013-01-01

77

Identification of a novel HLA-B*27 allele, B*27:79 and the B*27 subtype polymorphism in the Hunan ethnic Han population of China.  

UK PubMed Central (United Kingdom)

This article describes a novel HLA-B*27 allele, HLA-B*27:79, which was identified in a Hunan Han ethnic individual of China by a PCR sequence-based typing method. The new sequence has one nucleotide mutation at position 437(A?T) compared with the allele B*27:04:01. This nucleotide change causes an amino acid substitution from Aspartate (Asp) to Valine (Val) at codon 122. This is the first report of mutation at this position in the HLA-B locus. Then, we investigated the HLA-B*27 subtype polymorphism of the Hunan Han population, and the results showed that B*27:04, B*27:05 and B*27:06 are the predominant subtypes with the allele frequencies 0.97%, 0.26% and 0.10% respectively.

Xie Y; Wang S; Zuo Z; Zhang G; Cao L; Li T

2013-04-01

78

Rapid HLA-B27 screening with real-time TaqMan PCR: a clinical validation in the Dutch population.  

UK PubMed Central (United Kingdom)

BACKGROUND: Human leukocyte antigen B27 (HLA-B27) is strongly associated with ankylosing spondylitis. The B27 allele is present in 90% of patients with this disease, whereas it is present in only 9% of Caucasians. Molecular detection of HLA-B27 is traditionally based on allele specific amplification of exon 2 (Olerup method) or exon 3 (Dominguez method) by PCR, followed by gel analysis. METHODS: We developed a real-time TaqMan PCR based on the Dominguez method with a ?-Globin PCR as internal control. RESULTS: A total of 544 clinical samples were used to compare the real-time TaqMan PCR with the traditional Dominguez PCR, the traditional Olerup PCR and a commercial Olerup based HLA-B27 detection kit (Olerup SSPTM HLA-B27, GenoVision). While 542 samples gave concordant results, two samples showed discrepancies and were further analyzed. One sample that showed a discrepancy was negative with the traditional Olerup method and positive with the three other procedures. Sequencing analysis showed the presence of HLA-B*2712 in this sample. The other sample, positive with both Olerup based PCRs and negative with both Dominguez based methods, turned out to be positive for HLA-B*2707 by sequence analysis. CONCLUSIONS: With a correct result for 543 out of 544 samples (99.8%), we consider our real-time HLA-B27 PCR is a reliable method to detect HLA-B27 in the Dutch population, with reduced hands-on time and contamination risk compared to traditional PCR methods.

Roelandse-Koop EA; Buisman B; van Hannen EJ; van der Zee A; Kortlandt W; Hermans MH; van Houte AJ; van Rhee-Luderer R

2011-12-01

79

Association of HLA-B27 genetic polymorphisms with ankylosing spondylitis susceptibility worldwide: a meta-analysis.  

UK PubMed Central (United Kingdom)

OBJECTIVES: Many publications have evaluated the correlation between HLA-B27 polymorphisms and ankylosing spondylitis (AS), with conflicting results. We carried out this new meta-analysis in order to collect all the relevant studies to further clarify the association of HLA-B27 polymorphisms with AS susceptibility. METHODS: Relevant published data were retrieved through Medline, PubMed, Web of Science, CNKI, and the Chinese BioMedical Literature Database on disc. The statistical analysis was conducted using Review Manager Version 5.0 and STATA 11.0. From these data, the odds ratio (OR) with a 95 % confidence interval (95 % CI) was calculated. RESULTS: (1) A total of 38 studies, including 3,410 AS cases and 1,735 healthy controls, were collected in this meta-analysis. (2) Our results showed that B2704 was a risk factor but B2703, B2706, B2707, B2727, B2729, and B2747 may be protective factors for AS worldwide. (3) These subtypes, such as B2701, B2702, B2705, B2708-15, B2717-20, B2723-24, B2733, B2735, B2740, B2746, B2749, and B2767, showed no association with susceptibility to AS. There was a huge difference with previous reports for B2702 and B2705. (4) The B2702, B2704 and B2705 subtypes have existed high heterogeneity but no publication bias. CONCLUSIONS: This meta-analysis in our study suggested that B2704 might be a potential risk factor, however, B2703, B2706, and B2707 might be potential protective factors of AS, especially in Asia.

Yang T; Duan Z; Wu S; Liu S; Zeng Z; Li G; Wang S; Fan D; Ye D; Xu S; Zhang L; Pan F

2013-02-01

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Normal distribution of complement C3b receptor (CR1) numbers on erythrocytes in blood donors: low levels are associated with HLA-B27.  

Science.gov (United States)

Blood donors were examined for C3b receptor (CR1) levels on erythrocytes by a microtiter plate enzyme-linked immunosorbent assay and for HLA-A, -B, and -C antigens, C3 phenotypes and blood groups. The CR1 levels among 150 donors varied in the range from 24 to 130% (mean +/- 1 SD = 69.9 +/- 22.9) of an erythrocyte standard, and in accordance with a polygenic model of inheritance the distribution of CR1 was well approximated by a normal curve (chi-square goodness-of-fit test nonsignificant, p = 0.66). Low CR1 levels were associated with HLA-B27 (n = 11, p = 0.05), which was confirmed by investigation of further 20 HLA-B27-positive donors (p = 0.05). The CR1 levels were not associated with a certain C3 phenotype. Possibly, the blood group phenotype M-M + S-s+ is associated with high and the S antigen with low CR1 numbers. The relation between low CR1 numbers and HLA-B27 might be important for the understanding of the association between HLA-B27 and certain inflammatory rheumatological diseases. PMID:2945696

Thomsen, B S; Jacobsen, S E; Nielsen, H; Jakobsen, B K; Sørensen, H

1986-01-01

 
 
 
 
81

Normal distribution of complement C3b receptor (CR1) numbers on erythrocytes in blood donors: low levels are associated with HLA-B27.  

UK PubMed Central (United Kingdom)

Blood donors were examined for C3b receptor (CR1) levels on erythrocytes by a microtiter plate enzyme-linked immunosorbent assay and for HLA-A, -B, and -C antigens, C3 phenotypes and blood groups. The CR1 levels among 150 donors varied in the range from 24 to 130% (mean +/- 1 SD = 69.9 +/- 22.9) of an erythrocyte standard, and in accordance with a polygenic model of inheritance the distribution of CR1 was well approximated by a normal curve (chi-square goodness-of-fit test nonsignificant, p = 0.66). Low CR1 levels were associated with HLA-B27 (n = 11, p = 0.05), which was confirmed by investigation of further 20 HLA-B27-positive donors (p = 0.05). The CR1 levels were not associated with a certain C3 phenotype. Possibly, the blood group phenotype M-M + S-s+ is associated with high and the S antigen with low CR1 numbers. The relation between low CR1 numbers and HLA-B27 might be important for the understanding of the association between HLA-B27 and certain inflammatory rheumatological diseases.

Thomsen BS; Jacobsen SE; Nielsen H; Jakobsen BK; Sørensen H

1986-01-01

82

Complete sequence of HLA-B27 cDNA identified through the characterization of structural markers unique to the HLA-A, -B, and -C allelic series  

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Antigen HLA-B27 is a high-risk genetic factor with respect to a group of rheumatoid disorders, especially ankylosing spondylitis. A cDNA library was constructed from an autozygous B-cell line expressing HLA-B27, HLA-Cw1, and the previously cloned HLA-A2 antigen. Clones detected with an HLA probe were isolated and sorted into homology groups by differential hybridization and restriction maps. Nucleotide sequencing allowed the unambiguous assignment of cDNAs to HLA-A, -B, and -C loci. The HLA-B27 mRNA has the structure features and the codon variability typical of an HLA class I transcript but it specifies two uncommon amino acid replacements: a cysteine in position 67 and a serine in position 131. The latter substitution may have functional consequences, because it occurs in a conserved region and at a position invariably occupied by a species-specific arginine in humans and lysine in mice. The availability of the complete sequence of HLA-B27 and of the partial sequence of HLA-Cw1 allows the recognition of locus-specific sequence markers, particularly, but not exclusively, in the transmembrane and cytoplasmic domains.

Szoets, H.; Reithmueller, G.; Weiss, E.; Meo, T.

1986-03-01

83

Association of IL1R polymorphism with HLA-B27 positive in Iranian patients with ankylosing spondylitis.  

Science.gov (United States)

Ankylosing spondylitis (AS) is one of the most common causes of inflammatory arthritis, with an estimated prevalence of 0.1-0.9%. Genetic factors have been strongly implicated in its aetiology, and heritability as assessed by twin studies has been estimated to be >90%. HLA- B27 is almost essential for inheritance of AS; it is not merely sufficient for explaining the pattern of familial recurrence of the disease. This study's purpose is to investigate the association of ankylosing spondylitis with single-nucleotide polymorphisms (SNPs) in the IL-1 family: IL-1a (-889C/T) rs1800587, IL-1b (-511C/T) rs16944, IL-1b (+3962C/T) rs1143634, IL-1R (Pst-1 1970C/T) rs2234650 and IL-1RA (Mspa-1 11100C/T) rs315952. 99 unrelated Iranian AS patients and 217 healthy control subjects were selected. Cytokine typing was performed by the polymerase chain reaction with sequence-specific primers assay. The allele and genotype frequencies of the polymorphisms were determined: The IL1? rs1800587, IL1? rs16944 and IL1? rs1143634 were not significantly associated with AS. Genotype frequencies at IL1R rs2234650 differed between cases and controls (?(2)=8.85; p=0.01); the IL1R rs2234650 C/T and T/T genotypes were less common in AS patients than controls. The IL1R rs2234650 C/T genotype was inversely associated with AS comparing with the IL1R rs2234650 C/C genotype (OR=0.48; p=0.005). IL1R rs2234650 C/T genotype was less common in patients than controls (OR=0.37; p=0.02).Furthermore IL1R rs2234650 T allele was strongly associated with HLA-B2702 patients rather than HLA-B2705 but was not associated with HLA-B27 negative patients (OR=0.33; p=0.01). Polymorphisms of IL1? rs1800587, IL1? rs16944 and IL1? rs1143634 were not significantly associated with ankylosing spondylitis but inversely in this study IL1R rs2234650 was significantly associated and carriage of T allele in IL1R rs2234650 seems to be protective, while carriage of C allele result in two fold higher risk of developing AS. PMID:22285486

Mahmoudi, M; Amirzargar, A A; Jamshidi, A R; Farhadi, E; Noori, S; Avraee, M; Nazari, B; Nicknam, M H

2011-12-01

84

Association of IL1R polymorphism with HLA-B27 positive in Iranian patients with ankylosing spondylitis.  

UK PubMed Central (United Kingdom)

Ankylosing spondylitis (AS) is one of the most common causes of inflammatory arthritis, with an estimated prevalence of 0.1-0.9%. Genetic factors have been strongly implicated in its aetiology, and heritability as assessed by twin studies has been estimated to be >90%. HLA- B27 is almost essential for inheritance of AS; it is not merely sufficient for explaining the pattern of familial recurrence of the disease. This study's purpose is to investigate the association of ankylosing spondylitis with single-nucleotide polymorphisms (SNPs) in the IL-1 family: IL-1a (-889C/T) rs1800587, IL-1b (-511C/T) rs16944, IL-1b (+3962C/T) rs1143634, IL-1R (Pst-1 1970C/T) rs2234650 and IL-1RA (Mspa-1 11100C/T) rs315952. 99 unrelated Iranian AS patients and 217 healthy control subjects were selected. Cytokine typing was performed by the polymerase chain reaction with sequence-specific primers assay. The allele and genotype frequencies of the polymorphisms were determined: The IL1? rs1800587, IL1? rs16944 and IL1? rs1143634 were not significantly associated with AS. Genotype frequencies at IL1R rs2234650 differed between cases and controls (?(2)=8.85; p=0.01); the IL1R rs2234650 C/T and T/T genotypes were less common in AS patients than controls. The IL1R rs2234650 C/T genotype was inversely associated with AS comparing with the IL1R rs2234650 C/C genotype (OR=0.48; p=0.005). IL1R rs2234650 C/T genotype was less common in patients than controls (OR=0.37; p=0.02).Furthermore IL1R rs2234650 T allele was strongly associated with HLA-B2702 patients rather than HLA-B2705 but was not associated with HLA-B27 negative patients (OR=0.33; p=0.01). Polymorphisms of IL1? rs1800587, IL1? rs16944 and IL1? rs1143634 were not significantly associated with ankylosing spondylitis but inversely in this study IL1R rs2234650 was significantly associated and carriage of T allele in IL1R rs2234650 seems to be protective, while carriage of C allele result in two fold higher risk of developing AS.

Mahmoudi M; Amirzargar AA; Jamshidi AR; Farhadi E; Noori S; Avraee M; Nazari B; Nicknam MH

2011-12-01

85

Distress, depression and coping in HLA-B27-associated anterior uveitis with focus on gender differences.  

UK PubMed Central (United Kingdom)

BACKGROUND/AIMS: To evaluate depression, coping with disease and stress, and the subjective impression of distress and/or life events as triggers for recurrences in HLA-B27-associated anterior uveitis (B27-AU), with attention to gender-specific characteristics. METHODS: 171 patients with a history of B27-AU responded to a postal survey performed between January 2006 and April 2008 using standardised psychological questionnaires: Beck Depression Inventory, Freiburg Questionnaire on Coping with Illness, and Stress Coping Inventory. RESULTS: Patients with B27-AU differed from healthy controls showing more depressive symptoms (Beck Depression Inventory, 31.6%), applying characteristic disease coping as well as negative stress coping strategies. Female B27-AU patients tended to react with depression and male patients to use negative stress coping strategies. 57.9% of patients believed that psychological distress was a trigger for relapses, and 34.5% stated specific life events. Together, this group of patients achieved higher depression scores and used more negative disease and stress coping styles than patients without perception of distress. CONCLUSION: Patients with B27-AU patients exhibited significant psychopathology concerning depression and disease coping. Distress and life events were subjectively suspected to be a trigger. By imparting knowledge to the patients on probable development of depressive moods and the role of stress/life events as trigger for relapses, as well as offering behaviour therapy to optimise coping, may help patients to cope better with B27-AU.

Maca SM; Schiesser AW; Sobala A; Gruber K; Pakesch G; Prause C; Barisani-Asenbauer T

2011-05-01

86

Identification of immunogenic HLA-B*27:05 binding peptides of salmonella outer membrane protein in patients with reactive arthritis and undifferentiated spondyloarthropathy.  

UK PubMed Central (United Kingdom)

OBJECTIVE: Salmonella outer membrane proteins (OMP) are major immunogenic targets to synovial fluid lymphocytes of patients with reactive arthritis (ReA)/undifferentiated spondyloarthropathy (uSpA). Because these patients have genetic predisposition to HLA-B*27 and its subtype HLA-B*27:05, we sought to identify immunogenic HLA-B*27:05-binding salmonella OMP peptides in patients with ReA/uSpA. METHODS: A total of 125 HLA-B*27:05-binding salmonella OMP peptides identified using ProPred-I software were synthesized and grouped in 23 pools. The peptide pools, along with crude enteric bacterial lysates and salmonella OMP, were cultured with synovial fluid (SF) or peripheral blood mononuclear cells (PBMC) from 23 patients with ReA/uSpA, 10 with rheumatoid arthritis (RA), and 10 healthy individuals in 96-well culture plates. Proliferation was measured by tritiated thymidine uptake and interferon-? (IFN-?) levels in culture supernatant. Individual peptides from pools having significant responses were retested with cryopreserved cells. Immunogenic peptides thus identified were further tested in 5 additional new patients with ReA/uSpA by flow cytometry. A Basic Local Alignment Search Tool program was used to search for similar peptides from a protein bank of arthritogenic bacteria and human protein. RESULTS: Nineteen of 23 SFMC from ReA/uSpA showed a significant proliferative response to salmonella OMP, with minimal response of PBMC (1/10) from ReA/uSpA, SFMC from RA (1/10), or PBMC from controls (1/10). Nine salmonella OMP peptides, QRAEMLPTL, SRSGLNIAL, LRFLYAKSL, RLEGTWVKL, ARCIAPYAL, KLFLTTAAL, YRNSDFFGL, QRPAVRVKL, and YRVGPGDVL, were identified. Response to QRAEMLPTL was seen in 6/7 HLA-B*27:05-positive patients. All immunogenic peptides had sequence similarity with peptides from arthritogenic bacterial proteins, while 5 had similarity with peptides from human proteins. CONCLUSION: Nine novel immunogenic OMP peptides binding to HLA-B*27:05 were identified that showed sequence similarity with other arthritogenic bacteria.

Singh AK; Aggarwal A; Chaurasia S; Misra R

2013-02-01

87

HLA-B27  

Science.gov (United States)

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

88

Genetic study confirms association of HLA-DPA1(?)01:03 subtype with ankylosing spondylitis in HLA-B27-positive populations.  

UK PubMed Central (United Kingdom)

The association of human leukocyte antigen (HLA)-B27 with ankylosing spondylitis (AS) has been known for over 38 years. However, it is not the only gene associated with AS. The aim of this study was to confirm the association of HLA markers around HLA-DPA1/DPB1 region with AS in HLA-B27 positive populations. Five SNPs (rs422544, rs6914849, rs92777535, rs3128968 and rs2295119) from the HLA-DPA1/DPB1 region were genotyped in 340 individuals HLA-B27-positive from Portugal (137 AS patients and 203 healthy controls). Characterizations of HLA-DPA1/DPB1 alleles were also performed. rs422544 revealed a significant association with AS (P<0.05) and sliding windows (SW) analysis showed association of some groups of adjacent SNPs within HLA-DPA1/DPB1 region with AS (P<0.05). We also found association of the HLA-DPA1(?)01:03 allele with AS (P<0.05). This is the first study that confirms the association of HLA markers and haplotypes around HLA-DPA1 and HLA-DPB1 with AS.

Díaz-Peña R; Castro-Santos P; Aransay AM; Brüges-Armas J; Pimentel-Santos FM; López-Larrea C

2013-06-01

89

Antibody response to Klebsiella pneumoniae 60 kDa protein in familial and sporadic ankylosing spondylitis: role of HLA-B27 and characterization as a GroEL-like protein.  

UK PubMed Central (United Kingdom)

OBJECTIVE: To study the antibody response of HLA-B27+ patients with ankylosing spondylitis (AS) and their first degree relatives to the 60 kDa protein of Klebsiella pneumoniae and to characterize this protein. METHODS: Sera from 84 individuals were analyzed by ELISA to determine the titer of antibodies against the 60 kDa protein of K. pneumoniae. Subjects were divided into 3 categories: Group 1: 44 HLA-B27+ AS related individuals (35 patients, 9 healthy controls); Group 2: 28 healthy B27- AS related individuals; and Group 3: 12 healthy B27- non-AS related subjects. The 60 kDa protein of K. pneumoniae was induced at 45 degrees C and purified by electroelution from sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It was characterized as a GroEL-like heat shock protein (HSP). The recognition of GroEL-like protein was confirmed by immunoblot of 2 dimension electrophoresis. The response to GroEL-like protein from other bacteria and the response to lipopolysaccharide (LPS) was also analyzed by immunoblot. RESULTS: HLA-B27+ individuals (Group 1), independent of their disease status, showed a significant higher response to the 60 kDa protein of K. pneumoniae than HLA-B27- subjects from Groups 2 and 3 (p < 0.0001). This protein was characterized as a HSP of the GroEL family and designated HSP60Kp. The GroEL of other enterobacteria as well as that of Mycobacterium leprae were recognized by HLA-B27+ individuals by immunoblot, whereas HLA-B27- individuals did not. LPS was not recognized by HLA-B27 positive or negative subjects. CONCLUSION: These findings suggest a relationship between HLA-B27 and the response to a GroEL-like protein that could have implications in AS.

Cancino-Díaz ME; Pérez-Salazar JE; Domínguez-López L; Escobar-Gutiérrez A; Granados-Arreola J; Jiménez-Zamudio L; Burgos-Vargas R; García-Latorre E

1998-09-01

90

Immunodominance of HLA-B27-restricted HIV KK10-specific CD8(+) T-cells is not related to naive precursor frequency.  

UK PubMed Central (United Kingdom)

The factors that determine the immunodominance, efficacy and almost ubiquitous presence of CD8(+) T-cell responses to the HLA-B27-restricted HIV-1 p24 Gag-derived KK10 epitope remain to be fully elucidated. Here, we show that neither the precursor frequency nor the priming capacity of KK10-reactive CD8(+) T-cells within the naïve pool differ substantially in comparison to other specificities. These data implicate alternative mechanisms in the relative protection conferred by CD8(+) T-cell responses to this epitope.

Iglesias MC; Briceno O; Gostick E; Moris A; Meaudre C; Price DA; Ungeheuer MN; Saez-Cirion A; Mallone R; Appay V

2013-01-01

91

Optimizing the identification of patients with axial spondyloarthritis in primary care--the case for a two-step strategy combining the most relevant clinical items with HLA B27.  

UK PubMed Central (United Kingdom)

OBJECTIVES: The combination of clinical items suggestive of inflammatory back pain has proved useful for early identification of patients with axial SpA (axSpA) in primary care. However, whether HLA B27 contributes to that is unclear, and published recommendations have advised against it. In this study, we reanalysed data of that trial in relation to the HLA B27 results. METHODS: Consecutive patients <45 years old (n = 950) with back pain (BP) >2 months presenting to 143 orthopaedists were referred to 36 rheumatologists who made the diagnosis. The predictive value of HLA B27 (n = 298) alone and in combination, including modelling and a two-step strategy, was calculated. RESULTS: Among all patients (mean age 36 years, 52% female, median duration of BP 32 months), 107 had axSpA (36%). Using a simple model, HLA B27 alone performed better than all combinations of clinical items and adding it did not improve likelihood ratios (LRs). Using modelling, two-phase strategies were analysed. Additional items were only relevant in the HLA B27-negative group: improvement by movement, buttock pain and psoriasis. Combining this information revealed the presumably best strategy to predict axSpA in primary care: more than one of these items or HLA B27 need to be present (sensitivity 80.4%, specificity 75.4%, LR+ 3.27 and LR- 0.26). CONCLUSION: This is the first study to show that patients with axSpA are more reliably identified in primary care by a strategy that includes HLA B27. Because of the two-step approach, the test needs to be performed in only about half of patients with chronic BP.

Braun A; Gnann H; Saracbasi E; Grifka J; Kiltz U; Letschert K; Braun J

2013-08-01

92

Protective effect of an ERAP1 haplotype in ankylosing spondylitis: investigating non-MHC genes in HLA-B27-positive individuals.  

UK PubMed Central (United Kingdom)

Objective. The association of non-MHC genes with AS has been recently suggested. We aimed to investigate the association of the ERAP1, IL23R and TNFSF15 regions and the susceptibility to and protection from AS in HLA-B27-positive individuals.Methods. A total of 200 unrelated AS patients and 559 healthy unrelated subjects, all HLA-B27 positive, were tested. Twenty single nucleotide polymorphisms (SNPs) were investigated in and near IL23R (nine SNPs), in ERAP1 (five SNPs) and in TNFSF15 (six SNPs).Results. ERAP1 rs30187 [odds ratio (OR) = 1.5, P = 4.7 × 10(-3)] had the strongest association with AS susceptibility. A protective effect was found in three of the ERAP1 SNPs: rs17482078 (OR = 0.7, P = 2.8 × 10(-2)), rs10050860 (OR = 0.7, P = 2.3 × 10(-2)), rs2287987 (OR = 0.6, P = 1.3 × 10(-2)). The ERAP1 haplotype rs17482078/rs10050860/rs30187/rs2287987-CCTT showed an association with AS susceptibility (P = 6.8 × 10(-3)) and a protective effect was identified in rs17482078/rs10050860/rs30187/rs2287987-TTCC (P = 3.1 × 10(-2)). Significant association with AS susceptibility was found in one IL23R marker (rs1004819, P = 4.3 × 10(-2), OR = 1.3). No associations were observed in the TNFSF15 region.Conclusion. The identification of a new protection haplotype in ERAP1 and the lack of association of the TNFSF15 region can provide new insights into the understanding of the mechanisms underlying the susceptibility to and protection from AS.

Bettencourt BF; Rocha FL; Alves H; Amorim R; Caetano-Lopes J; Vieira-Sousa E; Pimentel-Santos F; Lima M; Porto G; Branco JC; Fonseca JE; Bruges-Armas J

2013-09-01

93

Can latent synergism of intestinal pathogens be responsible for inflammaging process causing Reiter's syndrome in a young patient HLA-B27 infected by atypical pathogens? A holistic view and clinical biochemical reinterpretation.  

Science.gov (United States)

A case of a genetically HLA-B27 patient fully investigated by molecular analyses, following a holistic vision and an anamnestic assessment of multi-site ecosystems is repeated. VDRL, Lupus anti-coagulant (LAC) and Widal-Wright (WWR), resulted positive. The antibodies (IgG/IgA anti-Ct) against chronic Chlamydia trachomatis inflammation were positive. In the context of all the enzymatic activities in reference range, the AMS and the ALP enzymatic activities showed an increasing trend and a time course augment depending respectively. Cultures, parasitological, digestibility tests and molecular analyses were then performed to investigate the different human ecosystems. Parasitological research and digestibility test were performed, resulting a latent chronic bowel inflammation, including certain enteroinvasive pathogens, such as, Salmonella, Shigella, Yersinia and Campylobacter (Enteric Pathogens Group, EPG) and Escherichia Coli pathogens (Escherichia Coli Pathogens Group, ECPG). The Salmonella typhi-DNA resulted positive, while 90% of the total microbic charge (TMC) was represented by C. freundi in culture analyses. Interpreting the VDRL positive test as early triggering of autoimmune disease, a few acute phase proteins as a pauci-symptomatic chronic phlogistic process, the amylase and alkaline phosphatase alterations as tissue markers of early intestinal inflammation, the Widal's reaction positivity together with the precocious clinical and faecal manifestations, this study suggests the prime triggering role of these atypical pathogens to cause a chronic low grade autoimmune response against the tissue/organ susceptible target, causing inflammaging phenomenon in young patient with chronic latent infection by Salmonella typhi, leading to Reiter's syndrome, in HLA-B27 positive patient. PMID:23241124

Del Boccio, M; Lobefalo, L; Pennelli, A; Toniato, E; Martinotti, S; Tenaglia, R; Neri, G; Del Boccio, G; Gallenga, P E

94

Can latent synergism of intestinal pathogens be responsible for inflammaging process causing Reiter's syndrome in a young patient HLA-B27 infected by atypical pathogens? A holistic view and clinical biochemical reinterpretation.  

UK PubMed Central (United Kingdom)

A case of a genetically HLA-B27 patient fully investigated by molecular analyses, following a holistic vision and an anamnestic assessment of multi-site ecosystems is repeated. VDRL, Lupus anti-coagulant (LAC) and Widal-Wright (WWR), resulted positive. The antibodies (IgG/IgA anti-Ct) against chronic Chlamydia trachomatis inflammation were positive. In the context of all the enzymatic activities in reference range, the AMS and the ALP enzymatic activities showed an increasing trend and a time course augment depending respectively. Cultures, parasitological, digestibility tests and molecular analyses were then performed to investigate the different human ecosystems. Parasitological research and digestibility test were performed, resulting a latent chronic bowel inflammation, including certain enteroinvasive pathogens, such as, Salmonella, Shigella, Yersinia and Campylobacter (Enteric Pathogens Group, EPG) and Escherichia Coli pathogens (Escherichia Coli Pathogens Group, ECPG). The Salmonella typhi-DNA resulted positive, while 90% of the total microbic charge (TMC) was represented by C. freundi in culture analyses. Interpreting the VDRL positive test as early triggering of autoimmune disease, a few acute phase proteins as a pauci-symptomatic chronic phlogistic process, the amylase and alkaline phosphatase alterations as tissue markers of early intestinal inflammation, the Widal's reaction positivity together with the precocious clinical and faecal manifestations, this study suggests the prime triggering role of these atypical pathogens to cause a chronic low grade autoimmune response against the tissue/organ susceptible target, causing inflammaging phenomenon in young patient with chronic latent infection by Salmonella typhi, leading to Reiter's syndrome, in HLA-B27 positive patient.

Del Boccio M; Lobefalo L; Pennelli A; Toniato E; Martinotti S; Tenaglia R; Neri G; Del Boccio G; Gallenga PE

2012-10-01

95

Transgenic Rat Models of Huntington's Disease.  

UK PubMed Central (United Kingdom)

Several animal models for Huntington's disease (HD) have been created in order to investigate mechanisms of disease, and to evaluate the potency of novel therapies. Here, we describe the characteristics of the two transgenic rat models: transgenic rat model of HD (fragment model) and the Bacterial Artificial Chromosome HD model (full-length model). We discuss their genetic, behavioural, neuropathological and neurophysiological features.

Carreira JC; Jahanshahi A; Zeef D; Kocabicak E; Vlamings R; von Hörsten S; Temel Y

2013-09-01

96

[HLA-B27 antigen and alkaptonuria  

UK PubMed Central (United Kingdom)

Study of urinary homogentisic acid and a determinantion of group HLA were carried out for 36 members of a family spread over three generations with three cases of ochronotic rheumatism in the second generation. Alkaptonuria was discovered in seven other subjects, six of them members of the third generation: urinary elimination was poor, less than 0.60 g/24 hours. There is a certain degree of consanguinity in the family studied here and these findings do not therefore rule out a recessive autosomal transmission of the alkaptonuria. They do however lead to the consideration that alkaptonuria may sometimes be found in heterozygotic subjects. A genetic relationship between HLA complex and alkaptonuria can only be claimed with difficulty from this familial study, but the high frequency of B 27 antigen (29 out of 36 members carring it) leaves room for the hypothesis that the B 27 gene, or more precisely a gene associated with the B 27 gene, plays a part in the development of ochronotic rheumatism.

Gaucher A; Netter P; Fuare G; Raffoux C; Chanson B; Baumgartner J; Psurel J; Streiff F

1977-04-01

97

Expression of a rat vasopressin transgene in rat testes.  

UK PubMed Central (United Kingdom)

The rat vasopressin gene contains two transcriptional promoters; the activity of one is confined to the hypothalamus, while the other is testis specific. To define the sequences mediating the cell-specific expression of the vasopressin gene, we introduced rat vasopressin transgenes into the rat germ line. Neither transgene 1.5-V beta gal-0.2, which consists of the entire vasopressin structural gene containing a 3 kbp beta-galactosidase reporter element in exon III, flanked by 1.5 kbp upstream of the start of hypothalamic transcription and 0.2 kbp downstream of the polyadenylation site, nor transgene 3-V beta gal-0.2, which consists of the entire VP structural gene containing a 3 kbp beta-galactosidase reporter element in exon III, flanked by 3 kbp upstream of the start of hypothalamic transcription and 0.2kbp downstream of the polyadenylation site, were expressed in the hypothalamus. This contrasts with a previously described transgene consisting of the rat vasopressin structural gene containing a reporter in exon III, flanked by 5 kbp of upstream and 3 kbp of downstream sequences, which is expressed in vasopressinergic hypothalamic neurons. Both the 3-V beta gal-0.2 and 1.5-V beta gal-0.2 transgenes were expressed in testicular germ cells using a promoter located within the beta-galactosidase reporter element. Transgene RNA was most abundant during the late stages of meiosis. Rats bearing vasopressin-beta-galactosidase transgenes provide new models for the study of the mechanisms whereby an epigenetic choice is made between the use of a germ cell or a somatic promoter, and the stage-specific transcriptional regulation of a germ cell promoter during spermatogenesis.

Zeng Q; Foo NC; Funkhouser JM; Carter DA; Murphy D

1994-11-01

98

Generation of Transgenic Rats through Induced Pluripotent Stem Cells.  

UK PubMed Central (United Kingdom)

The rat is an important animal model for human disease research. Using inhibitors of GSK3 and MAPK signaling pathways, rat embryonic stem cells and rat induced pluripotent stem cells (riPSCs) have been derived. However, unlike rat embryonic stem cells, germline competent riPSCs have only been derived from Wistar rats at low efficiency. Here, we found that an optimized induction medium containing knockout serum replacement (KOSR) and vitamin C (Vc) improved the rate and efficiency of riPSC generation from Dark Agouti rat fibroblasts and Sertoli cells. riPSCs maintained an undifferentiated status for more than 30 passages, and could differentiate into various cells types including germ cells when injected into rat blastocysts. Moreover, transgenic riPSCs could be generated through the PiggyBac transposon, which could be used to generate transgenic rats through germline transmission. riPSCs can be used as a novel tool in genetic and genomic studies of the rat.

Jiang MG; Li T; Feng C; Fu R; Yuan Y; Zhou Q; Li X; Wan H; Wang L; Li W; Xiao Y; Zhao XY; Zhou Q

2013-08-01

99

Conditional gene expression systems in the transgenic rat brain  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Turning gene expression on and off at will is one of the most powerful tools for the study of gene function in vivo. While several conditional systems were successful in invertebrates, in mice the Cre/loxP recombination system and the tet-controlled transcription activation system are predominant. Both expression systems allow for spatial and temporal control of gene activities, and, in the case of tet regulation, even for the reversible activation/inactivation of gene expression. Although the rat is the principal experimental model in biomedical research, in particular in studies of neuroscience, conditional rat transgenic systems are exceptionally rare in this species. Results We addressed this lack of technology, and established and thoroughly characterized CreERT2 and tTA transgenic rats with forebrain-specific transgene expression, controlled by the CaMKII alpha promoter. In addition, we developed new universal rat reporter lines for both transcription control systems and established inducible and efficient reporter gene expression in forebrain neurons. Conclusions We demonstrate that conditional genetic manipulations in the rat brain are both feasible and practicable and outline advantages and limitations of the Tet and Cre/loxP system in the rat brain.

Schönig Kai; Weber Tillmann; Frömmig Ariana; Wendler Lena; Pesold Brigitte; Djandji Dominik; Bujard Hermann; Bartsch Dusan

2012-01-01

100

Generation of Transgenic Rats through Induced Pluripotent Stem Cells.  

UK PubMed Central (United Kingdom)

The rat is an important animal model for human disease research. Using inhibitors of glycogen synthase kinase 3 and MAPK signaling pathways, rat embryonic stem cells and rat induced pluripotent stem cells (riPSCs) have been derived. However, unlike rat embryonic stem cells, germ line competent riPSCs have only been derived from Wistar rats at low efficiency. Here, we found that an optimized induction medium containing knock-out serum replacement and vitamin C improved the rate and efficiency of riPSCs generation from Dark Agouti rat fibroblasts and Sertoli cells. riPSCs maintained an undifferentiated status for >30 passages and could differentiate into various cells types including germ cells when injected into rat blastocysts. Moreover, transgenic riPSCs could be generated through the PiggyBac transposon, which could be used to generate transgenic rats through germ line transmission. riPSCs can be used as a novel tool in genetic and genomic studies of the rat.

Jiang MG; Li T; Feng C; Fu R; Yuan Y; Zhou Q; Li X; Wan H; Wang L; Li W; Xiao Y; Zhao XY; Zhou Q

2013-09-01

 
 
 
 
101

High blood pressure in transgenic mice carrying the rat angiotensinogen gene.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Transgenic mice were generated by injecting the entire rat angiotensinogen gene into the germline of NMRI mice. The resulting transgenic animals were characterized with respect to hemodynamics, parameters of the renin angiotension system, and expression of the transgene. The transgenic line TGM(rAOG...

Kimura, S; Mullins, J J; Bunnemann, B; Metzger, R; Hilgenfeldt, U; Zimmermann, F; Jacob, H; Fuxe, K; Ganten, D; Kaling, M

102

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; Frank Ocasio; Joseq A. Beltran

2007-01-01

103

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.

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

2008-01-01

104

Modeling Alzheimer's disease with non-transgenic rat models.  

UK PubMed Central (United Kingdom)

Alzheimer's disease (AD), for which there is no cure, is the most common form of dementia in the elderly. Despite tremendous efforts by the scientific community, the AD drug development pipeline remains extremely limited. Animal models of disease are a cornerstone of any drug development program and should be as relevant as possible to the disease, recapitulating the disease phenotype with high fidelity, to meaningfully contribute to the development of a successful therapeutic agent. Over the past two decades, transgenic models of AD based on the known genetic origins of familial AD have significantly contributed to our understanding of the molecular mechanisms involved in the onset and progression of the disease. These models were extensively used in AD drug development. The numerous reported failures of new treatments for AD in clinical trials indicate that the use of genetic models of AD may not represent the complete picture of AD in humans and that other types of animal models relevant to the sporadic form of the disease, which represents 95% of AD cases, should be developed. In this review, we will discuss the evolution of non-transgenic rat models of AD and how these models may open new avenues for drug development.

Lecanu L; Papadopoulos V

2013-05-01

105

Antinociceptive response in transgenic mice expressing rat tonin.  

Science.gov (United States)

Angiotensin II (Ang II) may be produced directly from angiotensinogen by tonin. Studies have demonstrated that Ang II and its metabolite Ang-(1-7) produce antinociception in pain animal models. The aim of the present study was to determine whether the transgenic mice that express rat tonin (TGM(rTon)) show altered nociceptive behavior and investigate the possible involvement of angiotensin metabolites. Nociception was evaluated using the thermal tail-flick and chemical acetic acid writhing tests, and the drugs were administered by intracerebroventricular and subcutaneous pathways, respectively. Probabilities less than 5% (Plosartan, an AT? receptor antagonist and A-779 (D-Ala7-Ang-(1-7)), a Mas receptor antagonist attenuated the antinociceptive behavior. Our data suggest that the Ang II produced in TGM(rTon) induces antinociception via the AT? receptor, while the Ang-(1-7) produced from Ang II induced antinociception via the Mas receptor. PMID:23665491

Pacheco, Daniela da Fonseca; Pacheco, Cinthia Mara da Fonseca; Lima, Mercia de Paula; Bader, Michael; Souza, Alexandro de Lima; Pesquero, Jorge Luiz; Castro Perez, Andrea; Duarte, Igor Dimitri Gama

2013-05-09

106

Functional recovery following peripheral nerve injury in the transgenic Thy1-GFP rat.  

UK PubMed Central (United Kingdom)

Transgenic mice have been previously used to assess nerve regeneration following peripheral nerve injury. However, mouse models are limited by their small caliber nerves, short nerve lengths, and their inability to fully participate during behavioral assessments. The transgenic Thy1 GFP rat is a novel transgenic rat model designed to assess regeneration following peripheral nerve injury. However, return of functional and behavioral recovery following nerve injury has not yet been evaluated in these rats. In this study, we ask whether differences in anatomy, recovery of locomotion, myological, and histomorphological measures exist between transgenic Thy1 GFP rats when compared to wild type (WT) Sprague Dawley rats following unilateral sciatic nerve injury. We found that both motor and sensory neuronal architecture, overground and skilled locomotion, muscle force, motor unit number estimation (MUNE) and wet muscle weights, and histomorphometric assessments are similar between both genetic phenotypes. Overall, these data support the use of the transgenic Thy1-GFP rat in experiments assessing functional and behavioral recovery following nerve injury and repair.

Kemp SW; Phua PD; Stanoulis KN; Wood MD; Liu EH; Gordon T; Borschel GH

2013-09-01

107

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 (L...

Martínez Navarrete, Gema Concepción; Seiler, M.J.; Aramant, R.B.; Fernández Sánchez, Laura; Pinilla Lozano, Isabel

108

Nonmammalian gonadotropin-releasing hormone molecules in the brain of promoter transgenic rats  

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Mammalian gonadotropin-releasing hormone (GnRH1) and nonmammalian immunoreactive GnRH subtypes were examined in transgenic rats carrying an enhanced GFP (EGFP) reporter gene driven by a rat GnRH1 promoter. Double-label immunocytochemistry was performed on EGFP+/GnRH1 brain sections by using antisera...

Parhar, Ishwar S.; Soga, Tomoko; Ogawa, Satoshi; Ogawa, Sonoko; Pfaff, Donald W.; Sakuma, Yasuo

109

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 injec...

Fernández Sánchez, Laura; Lax Zapata, Pedro; Pinilla Lozano, Isabel; Martín Nieto, José; Cuenca Navarro, Nicolás

110

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

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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.

Sheng Yi; Lin Chih-Cheng; Yue Junming; Sukhwani Meena; Shuttleworth Jennifer J; Chu Tianjiao; Orwig Kyle E

2010-01-01

111

Safety assessment of meat from transgenic cattle by 90-day feeding study in rats.  

Science.gov (United States)

The study was carried out to evaluate the subchronic toxicity of meat derived from human lactoferrin gene-modified cattle in male and female Wistar rats. Rats were fed 5% or 10% transgenic meat diet, 5% or 10% conventional meat diet, or AIN93G diet for 90 days. During the study, body weight and food consumption were weighed weekly and clinical observations were conducted daily. At the end of the study, urinary examination, hematology and blood biochemistry examination, macroscopic and microscopic examinations were performed. There were no biologically significant differences in these factors between the rat groups fed transgenic meat diet and conventional meat diet. Therefore, the present 90-day rodent feeding study suggests that meat derived from the transgenic cattle is equivalent to meat from conventional cattle in use as dietary supplements. PMID:23583492

Liu, Shan; Li, Chen-Xi; Feng, Xiao-Lian; Wang, Hui-Ling; Liu, Hai-Bo; Zhi, Yuan; Geng, Gui-Ying; Zhao, Jie; Xu, Hai-Bin

2013-04-10

112

Safety assessment of meat from transgenic cattle by 90-day feeding study in rats.  

UK PubMed Central (United Kingdom)

The study was carried out to evaluate the subchronic toxicity of meat derived from human lactoferrin gene-modified cattle in male and female Wistar rats. Rats were fed 5% or 10% transgenic meat diet, 5% or 10% conventional meat diet, or AIN93G diet for 90 days. During the study, body weight and food consumption were weighed weekly and clinical observations were conducted daily. At the end of the study, urinary examination, hematology and blood biochemistry examination, macroscopic and microscopic examinations were performed. There were no biologically significant differences in these factors between the rat groups fed transgenic meat diet and conventional meat diet. Therefore, the present 90-day rodent feeding study suggests that meat derived from the transgenic cattle is equivalent to meat from conventional cattle in use as dietary supplements.

Liu S; Li CX; Feng XL; Wang HL; Liu HB; Zhi Y; Geng GY; Zhao J; Xu HB

2013-07-01

113

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  

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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.

Agca Cansu; Fritz Jason J; Walker Lary C; Levey Allan I; Chan Anthony WS; Lah James J; Agca Yuksel

2008-01-01

114

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

115

Enhancement of tongue carcinogenesis in Hras128 transgenic rats treated with 4-nitroquinoline 1-oxide.  

Science.gov (United States)

Transgenic rats carrying human c-Ha-ras proto-oncogene (Hras128 rats) have been shown to be highly susceptible to induction of tumors. We have found an early induction of tongue tumors in Hras128 rats treated with 4-nitroquinoline 1-oxide (4NQO). 4NQO was administered to the Hras128 and wild-type Sprague-Dawley (SD) rats for 4 and 8 weeks, respectively. The experiment was terminated at 14 (Hras128 rats) and 28 (SD rats) weeks. Either during or after treatment with 4NQO, dysplastic hyperplasia, papilloma and squamous cell carcinoma were found on the tongue of both Hras128 and wild-type rats, with a higher incidence and multiplicity in Hras128 rats. Treatment of the Hras128 rats with 4NQO significantly increased cell proliferation in the tumor compared to the control rats. In the tongue tumors of the Hras128 rats, there was a significant increase in the mRNA expression levels of cyclin D1 and COX2. To examine whether this experimental system is useful for screening of the candidate agents for cancer preventive effect, nimesulide, a selective COX2 inhibitor, was tested in the present model. Nimesulide significantly decreased total multiplicity of tongue lesions compared to the control rats. Treatment of Hras128 rats with nimesulide caused a significant decrease in the levels of mRNA expression of cyclin D1 and COX2 in the tumor. Therefore, the current 4NQO-induced Hras128 rat tongue carcinogenesis model provides a simple and rapid system for investigating carcinogenesis process and evaluating the effect of possible cancer preventive agents for human tongue cancer. PMID:20043093

Naoi, Kuniko; Sunagawa, Nao; Yoshida, Ichiro; Morioka, Takamitsu; Nakashima, Makoto; Ishihara, Masashi; Fukamachi, Katsumi; Itoh, Yoshinori; Tsuda, Hiroyuki; Yoshimi, Naoki; Suzui, Masumi

2010-02-01

116

Enhancement of tongue carcinogenesis in Hras128 transgenic rats treated with 4-nitroquinoline 1-oxide.  

UK PubMed Central (United Kingdom)

Transgenic rats carrying human c-Ha-ras proto-oncogene (Hras128 rats) have been shown to be highly susceptible to induction of tumors. We have found an early induction of tongue tumors in Hras128 rats treated with 4-nitroquinoline 1-oxide (4NQO). 4NQO was administered to the Hras128 and wild-type Sprague-Dawley (SD) rats for 4 and 8 weeks, respectively. The experiment was terminated at 14 (Hras128 rats) and 28 (SD rats) weeks. Either during or after treatment with 4NQO, dysplastic hyperplasia, papilloma and squamous cell carcinoma were found on the tongue of both Hras128 and wild-type rats, with a higher incidence and multiplicity in Hras128 rats. Treatment of the Hras128 rats with 4NQO significantly increased cell proliferation in the tumor compared to the control rats. In the tongue tumors of the Hras128 rats, there was a significant increase in the mRNA expression levels of cyclin D1 and COX2. To examine whether this experimental system is useful for screening of the candidate agents for cancer preventive effect, nimesulide, a selective COX2 inhibitor, was tested in the present model. Nimesulide significantly decreased total multiplicity of tongue lesions compared to the control rats. Treatment of Hras128 rats with nimesulide caused a significant decrease in the levels of mRNA expression of cyclin D1 and COX2 in the tumor. Therefore, the current 4NQO-induced Hras128 rat tongue carcinogenesis model provides a simple and rapid system for investigating carcinogenesis process and evaluating the effect of possible cancer preventive agents for human tongue cancer.

Naoi K; Sunagawa N; Yoshida I; Morioka T; Nakashima M; Ishihara M; Fukamachi K; Itoh Y; Tsuda H; Yoshimi N; Suzui M

2010-02-01

117

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

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

Clary Caroline R; Guidot Daniel M; Bratina Margaux A; Otis Jeffrey S

2011-01-01

118

Hypertension is positively associated with prostate cancer development in the TRAP transgenic rat model.  

UK PubMed Central (United Kingdom)

Epidemiological data on the relationship between hypertension and prostate cancer development are conflicting. To cast light on this question, we performed animal experiments using hybrid rats generated by crossing the spontaneously hypertensive rat (SHR) or its normotensive control Wistar Kyoto (WKY) rat with a transgenic rat for adenocarcinoma of prostate (TRAP) that features development of adenocarcinoma at high incidence by 15 weeks of age. The number of adenocarcinomatous foci in the lateral prostate of hypertensive (TRAP × SHR)F1 rats was demonstrated to be significantly increased compared with those of normotensive (TRAP × WKY)F1 rats. In the ventral prostate, increase of carcinoma foci was also observed but did not reach significance. The number of cancer foci showing microinvasion in (TRAP × SHR)F1 rats was higher than that of (TRAP × WKY)F1 rats, but again without significance, while treatment with prazosin, an anti-hypertensive agent, tended to decrease microinvasive carcinoma foci in both the ventral and lateral prostate. In conclusion, the present study provided additional evidence that high blood pressure is associated with prostate cancer risk.

Takeshita K; Takahashi S; Tang M; Seeni A; Asamoto M; Shirai T

2011-04-01

119

Exogenous seeding of cerebral ?-amyloid deposition in ?APP-transgenic rats.  

UK PubMed Central (United Kingdom)

Deposition of the amyloid-? (A?) peptide in senile plaques and cerebral A? angiopathy (CAA) can be stimulated in A?-precursor protein (APP)-transgenic mice by the intracerebral injection of dilute brain extracts containing aggregated A? seeds. Growing evidence implicates a prion-like mechanism of corruptive protein templating in this phenomenon, in which aggregated A? itself is the seed. Unlike prion disease, which can be induced de novo in animals that are unlikely to spontaneously develop the disease, previous experiments with A? seeding have employed animal models that, as they age, eventually will generate A? lesions in the absence of seeding. In the present study, we first established that a transgenic rat model expressing human APP (APP21 line) does not manifest endogenous deposits of A? within the course of its median lifespan (30?months). Next, we injected 3-month-old APP21 rats intrahippocampally with dilute Alzheimer brain extracts containing aggregated A?. After a 9-month incubation period, these rats had developed senile plaques and CAA in the injected hippocampus, whereas control rats remained free of such lesions. These findings underscore the co-dependence of agent and host in governing seeded protein aggregation, and show that cerebral A?-amyloidosis can be induced even in animals that are relatively refractory to the spontaneous origination of parenchymal and vascular deposits of A?.

Rosen RF; Fritz JJ; Dooyema J; Cintron AF; Hamaguchi T; Lah JJ; LeVine H 3rd; Jucker M; Walker LC

2012-03-01

120

Proteomic analysis of kidneys from selenoprotein M transgenic rats in response to increased bioability of selenium  

Science.gov (United States)

Background To characterize changes in global protein expression in kidneys of transgenic rats overexpressing human selenoprotein M (SelM) in response to increased bioabivility of selenium (Sel), total proteins extracted from kidneys of 10-week-old CMV/hSelM Tg and wild-type rats were separated by 2-dimensional gel electrophoresis and measured for changes in expression. Results Ten and three proteins showing high antioxidant enzymatic activity were up- and down-regulated, respectively, in SelM-overexpressing CMV/hSelM Tg rats compared to controls based on an arbitrary 2-fold difference. Up-regulated proteins included LAP3, BAIAP2L1, CRP2, CD73 antigen, PDGF D, KIAA143 homolog, PRPPS-AP2, ZFP313, HSP-60, and N-WASP, whereas down-regulated proteins included ALKDH3, rMCP-3, and STC-1. After Sel treatment, five of the up-regulated proteins were significantly increased in expression in wild-type rats, whereas there were no changes in CMV/hSelM Tg rats. Only two of the down-regulated proteins showed reduced expression in wild-type and Tg rats after Sel treatment. Conclusions These results show the primary novel biological evidences that new functional protein groups and individual proteins in kidneys of Tg rats relate to Sel biology including the response to Sel treatment and SelM expression.

2013-01-01

 
 
 
 
121

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

Science.gov (United States)

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 test their resistance against complement (C)-mediated rejection by non-human primates. MATERIALS AND METHODS: Transgenic rats were generated by using a construct containing the human DAF cDNA under the transcriptional control of the endothelial cell (EC)-specific human ICAM-2 promoter. DAF expression was evaluated by immunohistology and by FACS analysis of purified ECs. Resistance of transgenic hearts against C-mediated damage was evaluated by ex vivo perfusion with human serum and by transplantation into cynomolgus monkeys. RESULTS: Immunohistological analysis of DAF expression in several organs from two transgenic lines showed uniform expression on the endothelium of all blood vessels. ECs purified from transgenic hearts showed 50% DAF expression compared to human ECs and >70% reduction of C-dependent cell lysis compared to control rat ECs. Hemizygous transgenic hearts perfused with human serum showed normal function for >60 min vs. 11. 2 +/- 1.7 min in controls. Hemi- or homozygous transgenic hearts transplanted into cynomolgus monkeys showed longer survival (15.2 +/- 7 min and >4.5 hr, respectively) than controls (5.5 +/- 1.4 min). In contrast to hyperacutely rejected control hearts, rejected homozygous DAF hearts showed signs of acute vascular rejection (AVR) characterized by edema, hemorrhage, and an intense PMN infiltration. CONCLUSIONS: We demonstrate that endothelial-specific DAF expression increased heart transplant survival in a rat-to-primate model of xenotransplantation. This will aid in the analysis of AVR and of new genes that may inhibit this form of rejection, thus helping to define strategies for the production of transgenic pigs. Images Fig. 1 Fig. 2 Fig. 3 Fig. 8

Charreau, B.; MA(C)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

122

Effect of HIV-1-related protein expression on cardiac and skeletal muscles from transgenic rats  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Human immunodeficiency virus type 1 (HIV-1) infection and the consequent acquired immunodeficiency syndrome (AIDS) has protean manifestations, including muscle wasting and cardiomyopathy, which contribute to its high morbidity. The pathogenesis of these myopathies remains partially understood, and may include nutritional deficiencies, biochemical abnormalities, inflammation, and other mechanisms due to viral infection and replication. Growing evidence has suggested that HIV-1-related proteins expressed by the host in response to viral infection, including Tat and gp120, may also be involved in the pathophysiology of AIDS, particularly in cells or tissues that are not directly infected with HIV-1. To explore the potentially independent effects of HIV-1-related proteins on heart and skeletal muscles, we used a transgenic rat model that expresses several HIV-1-related proteins (e.g., Tat, gp120, and Nef). Outcome measures included basic heart and skeletal muscle morphology, glutathione metabolism and oxidative stress, and gene expressions of atrogin-1, muscle ring finger protein-1 (MuRF-1) and Transforming Growth Factor-?1 (TGF?1), three factors associated with muscle catabolism. Results Consistent with HIV-1 associated myopathies in humans, HIV-1 transgenic rats had increased relative heart masses, decreased relative masses of soleus, plantaris and gastrocnemius muscles, and decreased total and myosin heavy chain type-specific plantaris muscle fiber areas. In both tissues, the levels of cystine (Cyss), the oxidized form of the anti-oxidant cysteine (Cys), and Cyss:Cys ratios were significantly elevated, and cardiac tissue from HIV-1 transgenic rats had altered glutathione metabolism, all reflective of significant oxidative stress. In HIV-1 transgenic rat hearts, MuRF-1 gene expression was increased. Further, HIV-1-related protein expression also increased atrogin-1 (~14- and ~3-fold) and TGF?1 (~5-fold and ~3-fold) in heart and plantaris muscle tissues, respectively. Conclusion We provide compelling experimental evidence that HIV-1-related proteins can lead to significant cardiac and skeletal muscle complications independently of viral infection or replication. Our data support the concept that HIV-1-related proteins are not merely disease markers, but rather have significant biological activity that may lead to increased oxidative stress, the stimulation of redox-sensitive pathways, and altered muscle morphologies. If correct, this pathophysiological scheme suggests that the use of dietary thiol supplements could reduce skeletal and cardiac muscle dysfunction in HIV-1-infected individuals.

Otis Jeffrey S; Ashikhmin Yaroslav I; Brown Lou; Guidot David M

2008-01-01

123

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

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

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

2012-01-01

124

In vivo genotoxicity of methyleugenol in gpt delta transgenic rats following medium-term exposure.  

UK PubMed Central (United Kingdom)

Methyleugenol (MEG), which is commonly used as a fragrance and flavoring agent, has been shown to induce hepatocellular tumors in rodents. However, the role of genotoxicity as a possible mechanism of action is not fully understood even though the DNA-reactive metabolite of MEG has been identified. In this study, a gpt delta transgenic rat model was used to clarify whether genotoxic mechanisms are involved in MEG-induced hepatocarcinogenesis following medium-term exposure. F344 gpt delta rats were subjected to repeated oral administration of MEG at dosages of 0, 10, 30, or 100mg/kg (a carcinogenic dose) for 13 weeks. The relative weight of the liver of the male and female rats that were administered 100mg/kg MEG and the absolute weight of the liver of the male rats that were administered 100mg/kg MEG were significantly increased. In addition, the number and area of glutathione S-transferase placental form (GST-P) positive foci and proliferating cell nuclear antigen (PCNA) positive cell ratios in the hepatocytes were significantly increased in the male and female rats that were administered 100mg/kg MEG compared with the control animals. In the in vivo mutation assays, a significant increase in the gpt and Spi(-) mutant frequencies was observed in both sexes at the carcinogenic dose. These results suggest the possible participation of genotoxic mechanisms in MEG-induced hepatocarcinogenesis.

Jin M; Kijima A; Hibi D; Ishii Y; Takasu S; Matsushita K; Kuroda K; Nohmi T; Nishikawa A; Umemura T

2013-02-01

125

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

126

Cellular immune response to Klebsiella pneumoniae antigens in patients with HLA-B27+ ankylosing spondylitis.  

UK PubMed Central (United Kingdom)

OBJECTIVE: To study the reactivity of peripheral blood mononuclear cells (PBMC) of patients with ankylosing spondylitis (AS) and rheumatoid arthritis (RA) and healthy controls to Klebsiella pneumoniae antigens and to the GroEL-like proteins from K. pneumoniae (HSP60Kp) and Mycobacterium leprae recombinant heat shock protein 65 (rHSP65Ml). METHODS: PBMC of 13 patients with AS and 9 with RA and 10 controls were stimulated in vitro by heat shock induced K. pneumoniae antigens in a cell blot assay, by insolubilized HSP60Kp, by cytosolic proteins (CP) from K. pneumoniae cultivated at 37 degrees C or 45 degrees C, by soluble HSP60Kp, or by rHSP65Ml. RESULTS: In the cell blot assay 7/13 AS and 3/9 RA patients responded to fraction 4, which contains mainly HSP60Kp, and no controls responded (AS vs. controls: p = 0.007). The response to the insolubilized HSP60Kp was positive in 6/13 AS patients but negative in RA patients and controls (p = 0.004). The response to CP45 degrees C was positive in 7/13 AS, in 2/9 RA, and no controls (AS vs controls: p<0.015). Response to the soluble HSP60Kp was found in 7/13 AS and 5/9 RA patients, but no controls (AS vs. controls: p = 0.0075). Response to rHSP65Ml was positive in 3/13 AS, 7/9 RA patients, and 1/10 controls (AS vs RA: p = 0.027; RA vs. controls: p = 0.005; AS vs. controls: nonsignificant). CONCLUSION: In PBMC of the majority of patients with AS and in some with RA, but not in healthy controls, there are cells that proliferate in the presence of HSP60 of K. pneumoniae.

Domínguez-López ML; Cancino-Díaz ME; Jiménez-Zamudio L; Granados-Arreola J; Burgos-Vargas R; García-Latorre E

2000-06-01

127

Role of Chlamydia trachomatis and HLA-B27 in sexually acquired reactive arthritis.  

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Inflammatory arthritis, tendinitis, and fasciitis after non-specific urethritis ("sexually acquired reactive arthritis" (SARA)) was studied prospectively in 531 men with non-specific urethritis, with particular reference to the frequency of isolation of Chlamydia trachomatis and the presence of HLA-...

Keat, A C; Maini, R N; Nkwazi, G C; Pegrum, G D; Ridgway, G L; Scott, J T

128

HLA-B27-Associated Reactive Arthritis: Pathogenetic and Clinical Considerations  

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Current evidence supports the concept that reactive arthritis (ReA) is an immune-mediated synovitis resulting from slow bacterial infections and showing intra-articular persistence of viable, nonculturable bacteria and/or immunogenetic bacterial antigens synthesized by metabolically active bacteria ...

Colmegna, Inés; Cuchacovich, Raquel; Espinoza, Luis R.

129

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

130

HLA-B27 and HLA-A2 subtypes: structure, evolution and function.  

UK PubMed Central (United Kingdom)

Beyond the resolution of tissue typing serology, HLA class I antigens display a certain level of structural microheterogeneity, that allows their subdivision into subtypes. The structure of these subtypes shows that multiple mechanisms operate in the generation of HLA polymorphism and suggests possible evolutionary pathways for subtype diversification. In addition, subtype polymorphism critically affects cellular allorecognition and antigen presentation to self-restricted T cells. These properties are used to define the structure and diversity of T-cell epitopes. In this review, José López de Castro discusses the nature and evolution of this polymorphism and its modulation of antigen recognition by cytolytic T lymphocytes.

López de Castro JA

1989-07-01

131

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

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

Theuer Juergen; Dechend Ralf; Muller Dominik N; Park Joon-Keun; Fiebeler Anette; Barta Peter; Ganten Detlev; Haller Hermann; Dietz Rainer; Luft Friedrich C

2002-01-01

132

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

UK PubMed Central (United Kingdom)

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

Nuber S; Harmuth F; Kohl Z; Adame A; Trejo M; Schönig K; Zimmermann F; Bauer C; Casadei N; Giel C; Calaminus C; Pichler BJ; Jensen PH; Müller CP; Amato D; Kornhuber J; Teismann P; Yamakado H; Takahashi R; Winkler J; Masliah E; Riess O

2013-02-01

133

Tauroursodeoxycholic acid prevents retinal degeneration in transgenic P23H rats.  

UK PubMed Central (United Kingdom)

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) recording. The effects of TUDCA on the number, morphology, integrity, and synaptic connectivity of retinal cells were characterized by immunofluorescence confocal microscopy. RESULTS: The amplitude of ERG a- and b-waves was significantly higher in TUDCA-treated animals under both scotopic and photopic conditions than in control animals. In the central area of the retina, TUDCA-treated P23H rats showed threefold more photoreceptors than control animals. The number of TUNEL-positive cells was significantly smaller in TUDCA-treated rats, in which photoreceptor morphology was preserved. Presynaptic and postsynaptic elements, as well as the synaptic contacts between photoreceptors and bipolar or horizontal cells, were preserved in TUDCA-treated P23H rats. Furthermore, in TUDCA-treated rat retinas, the number of both rod bipolar and horizontal cell bodies, as well as the density of their synaptic terminals in the outer plexiform layer, was greater than in control rats. CONCLUSIONS: TUDCA treatment was capable of preserving cone and rod structure and function, together with their contacts with their postsynaptic neurons. The neuroprotective effects of TUDCA make this compound potentially useful for delaying retinal degeneration in RP.

Fernández-Sánchez L; Lax P; Pinilla I; Martín-Nieto J; Cuenca N

2011-07-01

134

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

Science.gov (United States)

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.

Faure, Alexis; Es-seddiqi, Mouna; Brown, Bruce L.; Nguyen, Hoa P.; Riess, Olaf; von Horsten, Stephan; Le Blanc, Pascale; Desvignes, Nathalie; Bozon, Bruno; El Massioui, Nicole; Doyere, Valerie

2013-01-01

135

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

Science.gov (United States)

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. PMID:24133419

Faure, Alexis; Es-Seddiqi, Mouna; Brown, Bruce L; Nguyen, Hoa P; Riess, Olaf; von Hörsten, Stephan; Le Blanc, Pascale; Desvignes, Nathalie; Bozon, Bruno; El Massioui, Nicole; Doyère, Valérie

2013-09-26

136

Transgenic rescue of defective Cd36 enhances myocardial adenylyl cyclase signaling in spontaneously hypertensive rats.  

Science.gov (United States)

Dysfunction or abnormalities in the regulation of fatty acid translocase Cd36, a multifunctional membrane protein participating in uptake of long-chain fatty acids, has been linked to the development of heart diseases both in animals and humans. We have previously shown that the Cd36 transgenic spontaneously hypertensive rat (SHR-Cd36), with a wild type Cd36, has higher susceptibility to ischemic ventricular arrhythmias when compared to spontaneously hypertensive rat (SHR) carrying a mutant Cd36 gene, which may have been related to increased ?-adrenergic responsiveness of these animals (Neckar et al., 2012 Physiol. Genomics 44:173-182). The present study aimed to determine whether the insertion of the wild type Cd36 into SHR would affect the function of myocardial G protein-regulated adenylyl cyclase (AC) signaling. ?-Adrenergic receptors (?-ARs) were characterized by radioligand-binding experiments and the expression of selected G protein subunits, AC, and protein kinase A (PKA) was determined by RT-PCR and Western blot analyses. There was no significant difference in the amount of trimeric G proteins, but the number of ?-ARs was higher (by about 35 %) in myocardial preparations from SHR-Cd36 as compared to SHR. Besides that, transgenic rats expressed increased amount (by about 20 %) of the dominant myocardial isoforms AC5/6 and contained higher levels of both nonphosphorylated (by 11 %) and phosphorylated (by 45 %) PKA. Differently stimulated AC activity in SHR-Cd36 significantly exceeded (by about 18-30 %) the enzyme activity in SHR. Changes at the molecular level were reflected by higher contractile responses to stimulation by the adrenergic agonist dobutamine. In summary, it can be concluded that the increased susceptibility to ischemic arrhythmias of SHR-Cd36 is attributable to upregulation of some components of the ?-AR signaling pathway, which leads to enhanced sensitization of AC and increased cardiac adrenergic responsiveness. PMID:23636771

Klevstig, Martina; Manakov, Dmitry; Kasparova, Dita; Brabcova, Iveta; Papousek, Frantisek; Zurmanova, Jitka; Zidek, Vaclav; Silhavy, Jan; Neckar, Jan; Pravenec, Michal; Kolar, Frantisek; Novakova, Olga; Novotny, Jiri

2013-05-01

137

Tissue-specific and hormonal regulation of the gene for rat prostatic steroid-binding protein in transgenic mice.  

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We investigated the tissue-specific and hormonal regulation of the gene for rat prostatic steroid-binding protein by introducing the C3(1) gene with 4-kilobase (kb) upstream and 2-kb downstream flanking sequences into transgenic mice. There was selective expression in the ventral prostate that was s...

Allison, J; Zhang, Y L; Parker, M G

138

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; Kim Hyung-Wook; Kellom Matthew; Greenstein Dede; Chen Mei; Kraft Andrew; Harry Gaylia; Rapoport Stanley; Basselin Mireille

2012-01-01

139

Endotoxin-induced cytokine and chemokine expression in the HIV-1 transgenic rat  

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Full Text Available Abstract Background Repeated exposure to a low dose of a bacterial endotoxin such as lipopolysaccharide (LPS) causes immune cells to become refractory to a subsequent endotoxin challenge, a phenomenon known as endotoxin tolerance (ET). During ET, there is an imbalance in pro- and anti-inflammatory cytokine and chemokine production, leading to a dysregulated immune response. HIV-1 viral proteins are known to have an adverse effect on the immune system. However, the effects of HIV-1 viral proteins during ET have not been investigated. Methods In this study, HIV-1 transgenic (HIV-1Tg) rats and control F344 rats (n = 12 ea) were randomly treated with 2 non-pyrogenic doses of LPS (LL) to induce ET, or saline (SS), followed by a high challenge dose of LPS (LL+L, SS+L) or saline (LL+S, SS+S). The gene expression of 84 cytokines, chemokines, and their receptors in the brain and spleen was examined by relative quantitative PCR using a PCR array, and protein levels in the brain, spleen, and serum of 7 of these 84 genes was determined using an electrochemiluminescent assay. Results In the spleen, there was an increase in key pro-inflammatory (IL1?, IL-1?, IFN-?) and anti-inflammatory (IL-10) cytokines, and inflammatory chemokines (Ccl2, Ccl7, and Ccl9,) in response to LPS in the SS+L and LL+L (ET) groups of both the HIV-1Tg and F344 rats, but was greater in the HIV-1Tg rats than in the F344. In the ET HIV-1Tg and F344 (LL+L) rats in the spleen, the LPS-induced increase in pro-inflammatory cytokines was diminished and that of the anti-inflammatory cytokine was enhanced compared to the SS+L group rats. In the brain, IL-1?, as well as the Ccl2, Ccl3, and Ccl7 chemokines were increased to a greater extent in the HIV-1Tg rats compared to the F344; whereas Cxcl1, Cxcl10, and Cxcl11 were increased to a greater extent in the F344 rats compared to the HIV-1Tg rats in the LL+L and SS+L groups. Conclusion Our data indicate that the continuous presence of HIV-1 viral proteins can have tissue-dependent effects on endotoxin-induced cytokine and chemokine expression in the ET state.

Homji Natasha F; Mao Xin; Langsdorf Erik F; Chang Sulie L

2012-01-01

140

Endotoxin-induced cytokine and chemokine expression in the HIV-1 transgenic rat.  

UK PubMed Central (United Kingdom)

BACKGROUND: Repeated exposure to a low dose of a bacterial endotoxin such as lipopolysaccharide (LPS) causes immune cells to become refractory to a subsequent endotoxin challenge, a phenomenon known as endotoxin tolerance (ET). During ET, there is an imbalance in pro- and anti-inflammatory cytokine and chemokine production, leading to a dysregulated immune response. HIV-1 viral proteins are known to have an adverse effect on the immune system. However, the effects of HIV-1 viral proteins during ET have not been investigated. METHODS: In this study, HIV-1 transgenic (HIV-1Tg) rats and control F344 rats (n = 12 ea) were randomly treated with 2 non-pyrogenic doses of LPS (LL) to induce ET, or saline (SS), followed by a high challenge dose of LPS (LL+L, SS+L) or saline (LL+S, SS+S). The gene expression of 84 cytokines, chemokines, and their receptors in the brain and spleen was examined by relative quantitative PCR using a PCR array, and protein levels in the brain, spleen, and serum of 7 of these 84 genes was determined using an electrochemiluminescent assay. RESULTS: In the spleen, there was an increase in key pro-inflammatory (IL1?, IL-1?, IFN-?) and anti-inflammatory (IL-10) cytokines, and inflammatory chemokines (Ccl2, Ccl7, and Ccl9,) in response to LPS in the SS+L and LL+L (ET) groups of both the HIV-1Tg and F344 rats, but was greater in the HIV-1Tg rats than in the F344. In the ET HIV-1Tg and F344 (LL+L) rats in the spleen, the LPS-induced increase in pro-inflammatory cytokines was diminished and that of the anti-inflammatory cytokine was enhanced compared to the SS+L group rats. In the brain, IL-1?, as well as the Ccl2, Ccl3, and Ccl7 chemokines were increased to a greater extent in the HIV-1Tg rats compared to the F344; whereas Cxcl1, Cxcl10, and Cxcl11 were increased to a greater extent in the F344 rats compared to the HIV-1Tg rats in the LL+L and SS+L groups. CONCLUSION: Our data indicate that the continuous presence of HIV-1 viral proteins can have tissue-dependent effects on endotoxin-induced cytokine and chemokine expression in the ET state.

Homji NF; Mao X; Langsdorf EF; Chang SL

2012-01-01

 
 
 
 
141

Transcriptome sequencing of gene expression in the brain of the HIV-1 transgenic rat.  

Science.gov (United States)

The noninfectious HIV-1 transgenic (HIV-1Tg) rat was developed as a model of AIDs-related pathology and immune dysfunction by manipulation of a noninfectious HIV-1(gag-pol) virus with a deleted 3-kb SphI-MscI fragment containing the 3' -region of gag and the 5' region of pol into F344 rats. Our previous studies revealed significant behavioral differences between HIV-1Tg and F344 control rats in their performance in the Morris water maze and responses to psychostimulants. However, the molecular mechanisms underlying these behavioral differences remain largely unknown. The primary goal of this study was to identify differentially expressed genes and enriched pathways affected by the gag-pol-deleted HIV-1 genome. Using RNA deep sequencing, we sequenced RNA transcripts in the prefrontal cortex, hippocampus, and striatum of HIV-1Tg and F344 rats. A total of 72 RNA samples were analyzed (i.e., 12 animals per group × 2 strains × 3 brain regions). Following deep-sequencing analysis of 50-bp paired-end reads of RNA-Seq, we used Bowtie/Tophat/Cufflinks suites to align these reads into transcripts based on the Rn4 rat reference genome and to measure the relative abundance of each transcript. Statistical analyses on each brain region in the two strains revealed that immune response- and neurotransmission-related pathways were altered in the HIV-1Tg rats, with brain region differences. Other neuronal survival-related pathways, including those encoding myelin proteins, growth factors, and translation regulators, were altered in the HIV-1Tg rats in a brain region-dependent manner. This study is the first deep-sequencing analysis of RNA transcripts associated the HIV-1Tg rat. Considering the functions of the pathways and brain regions examined in this study, our findings of abnormal gene expression patterns in HIV-1Tg rats suggest mechanisms underlying the deficits in learning and memory and vulnerability to drug addiction and other psychiatric disorders observed in HIV-positive patients. PMID:23536882

Li, Ming D; Cao, Junran; Wang, Shaolin; Wang, Ju; Sarkar, Sraboni; Vigorito, Michael; Ma, Jennie Z; Chang, Sulie L

2013-03-25

142

Transcriptome sequencing of gene expression in the brain of the HIV-1 transgenic rat.  

UK PubMed Central (United Kingdom)

The noninfectious HIV-1 transgenic (HIV-1Tg) rat was developed as a model of AIDs-related pathology and immune dysfunction by manipulation of a noninfectious HIV-1(gag-pol) virus with a deleted 3-kb SphI-MscI fragment containing the 3' -region of gag and the 5' region of pol into F344 rats. Our previous studies revealed significant behavioral differences between HIV-1Tg and F344 control rats in their performance in the Morris water maze and responses to psychostimulants. However, the molecular mechanisms underlying these behavioral differences remain largely unknown. The primary goal of this study was to identify differentially expressed genes and enriched pathways affected by the gag-pol-deleted HIV-1 genome. Using RNA deep sequencing, we sequenced RNA transcripts in the prefrontal cortex, hippocampus, and striatum of HIV-1Tg and F344 rats. A total of 72 RNA samples were analyzed (i.e., 12 animals per group × 2 strains × 3 brain regions). Following deep-sequencing analysis of 50-bp paired-end reads of RNA-Seq, we used Bowtie/Tophat/Cufflinks suites to align these reads into transcripts based on the Rn4 rat reference genome and to measure the relative abundance of each transcript. Statistical analyses on each brain region in the two strains revealed that immune response- and neurotransmission-related pathways were altered in the HIV-1Tg rats, with brain region differences. Other neuronal survival-related pathways, including those encoding myelin proteins, growth factors, and translation regulators, were altered in the HIV-1Tg rats in a brain region-dependent manner. This study is the first deep-sequencing analysis of RNA transcripts associated the HIV-1Tg rat. Considering the functions of the pathways and brain regions examined in this study, our findings of abnormal gene expression patterns in HIV-1Tg rats suggest mechanisms underlying the deficits in learning and memory and vulnerability to drug addiction and other psychiatric disorders observed in HIV-positive patients.

Li MD; Cao J; Wang S; Wang J; Sarkar S; Vigorito M; Ma JZ; Chang SL

2013-01-01

143

Angiotensin-(1-7) attenuates the anxiety and depression-like behaviors in transgenic rats with low brain angiotensinogen.  

UK PubMed Central (United Kingdom)

Transgenic rats with low brain angiotensinogen, TGR(ASrAOGEN)680, expressing an antisense RNA against angiotensinogen in glial cells, provide an interesting tool to evaluate the role of brain angiotensins in different behavior responses. The present study was conducted to test the hypothesis that angiotensin-(1-7) [Ang-(1-7)] and serotonin can modulate anxiety and depression-related behaviors in the TGR(ASrAOGEN)680 rats. Therefore, the effect of acute intracerebroventricular administration of Ang-(1-7) and intraperitoneal administration of the selective serotonin reuptake inhibitor fluoxetine was evaluated in TGR(ASrAOGEN) rats subjected to an elevated plus maze (EPM) and forced swimming (FST) tests. Transgenic rats spent a lower percentage of time in the open arms of EPM and showed a significant increase in the immobility time in FST, indicating that a low angiotensinogen level in the brain leads to anxiety-like behavior accompanied by a depression-like state. Administration of both, Ang-(1-7) and fluoxetine reversed the anxiety- and depressive-like behavior of transgenic rats with low brain angiotensinogen, suggesting that this may be, at least in part, related to a decreased level of Ang-(1-7) and serotonin in the brain of these animals.

Kangussu LM; Almeida-Santos AF; Bader M; Alenina N; Fontes MA; Santos RA; Aguiar DC; Campagnole-Santos MJ

2013-09-01

144

Derivation and characterization of embryonic stem cells lines derived from transgenic Fischer 344 and Dark Agouti rats.  

UK PubMed Central (United Kingdom)

Rat embryonic stem cell (ESC) lines are not widely available, and there are only 2 lines available for distribution. Here, ESC lines were derived and characterized from Fischer 344 (F344) rats that express marker transgenes either ?-galactosidase or human placental alkaline phosphatase (AP), nontransgenic F344 rats, and from Dark Agouti (DA) rats. The ESC lines were maintained in an undifferentiated state as characterized by colony morphology, expression of Oct4, Nanog, Sox-2, Cdx2, and Stella, staining for AP, and stage-specific embryonic antigen-1. Pluripotency was demonstrated in vitro by differentiation to embryoid bodies, followed by embryonic monsters. The Cdx2 expression by ESCs was unexpected and was confirmed via reverse transcriptase-polymerase chain reaction, immunocytochemistry. Pluripotency of ESCs was demonstrated in vivo by production of teratoma after an injection into F344 nontransgenic rats, and by an injection of male DA ESCs into F344 or Sprague-Dawley rat blastocysts and the generation of chimeric rats and germline contribution. ESCs from both F344 and DA contributed to chimeric rats, and one DA ESC line was proved to be germline competent. ESC sublines were created by transfection with a plasmid expressing enhanced green fluorescent protein (eGFP) under the control of a beta actin promoter and cytomegalovirus enhancer (pCX-eGFP) or by transfection with a plasmid expressing GFP under the control of a 3.1 kb portion of the rat Oct4 promoter (pN1-Oct4-GFP). In pN1-Oct4-GFP sublines, GFP gene expression and fluorescence were shown to be correlated with endogenous Oct4 gene expression. Therefore, these new ESC lines may be useful for tissue engineering and transplantation studies or for optimizing culture conditions required for self-renewal and differentiation of rat ESCs. While they made chimeric rats, further work is needed to confirm whether the transgenic F344 rat ESCs described here are germline-competent ESCs.

Hong J; He H; Weiss ML

2012-06-01

145

Neurobehavioral alterations in HIV-1 transgenic rats: evidence for dopaminergic dysfunction.  

UK PubMed Central (United Kingdom)

Clinical studies have provided evidence that the progression of HIV-1-associated neurocognitive disorders (HAND) involves alterations in dopamine (DA) systems. Drugs of abuse that act on the brain DA system, such as cocaine (Coc), may exacerbate HIV-1 infection and consequent behavioral and neurological manifestations. In the present study, we used the HIV-1 transgenic (Tg) rat, which constitutively expresses 7 of the 9 HIV-1 genes, to assess potential DA system alterations in three behavioral assays: prepulse inhibition (PPI) of the auditory startle response (ASR), novelty and habituation/retention, and sensitization to Coc across repeated administration. Adult female Sprague-Dawley rats were tested in each experiment. The HIV-1 Tg animals were hyperreactive to auditory startle stimuli and displayed a leftward shift in the temporal window for maximal PPI, suggesting an alteration in sensorimotor gating. All animals displayed an initial robust locomotor response to a novel environment which dissipated with repeated testing; however, the HIV-1 Tg rats, relative to controls, consistently showed a weaker novelty response across monthly-spaced assessments. The HIV-1 Tg animals also showed decreased intrasession habituation of motor activity across 3-day periods that emerged across monthly-spaced locomotor activity sessions; a pattern consistent with impaired long-term episodic memory. Furthermore, the HIV-1 Tg group displayed differential cocaine-induced sensitization, observed both in initiation across the 10-day cocaine treatment, and in expression following a cocaine rechallenge after a 7-day abstinence. Collectively, the present data implicate that the non-infectious HIV-1 Tg rat, which resembles the complete suppression of infection in HIV-1 positive individuals under CART, displays sustained, if not permanent, alterations in the brain DA system.

Moran LM; Booze RM; Webb KM; Mactutus CF

2013-01-01

146

Early deficits in declarative and procedural memory dependent behavioral function in a transgenic rat model of Huntington's disease.  

UK PubMed Central (United Kingdom)

In Huntington's disease (HD) cognitive deficits co-exist with motor impairments, both contributing to the overall disease symptomology. Despite short-term and working memory impairments, learning and other non-motoric behavioral deficits arising from the damage to frontostriatal loop being common in HD patients, most of the experimental work with transgenic animals focuses on motor symptoms. The transgenic rat model (tgHD) recapitulates many hallmark HD-like symptoms, such as huntingtin aggregates, cellular loss and dysfunction, and motor, and some cognitive deficits. In the current study we tested tgHD rats in two different cognitive, water maze competition paradigms to learn more about the impact of the transgene on learning and memory processing using hippocampal- and striatal-based memory systems. The tgHD rats had early and robust cognitive deficits in learning and memory function in both paradigms. Specifically, the transgenic animals were impaired in task acquisition and committed more procedural errors with the strongest phenotype amongst the homozygote tgHD. Although the transgenic animals were capable of using both procedural and declarative memory, their response patterns were distinct from wild-type animals. Wide spread huntingtin aggregates were observed at 13 months, but neither PET nor autoradiography indicated neuronal loss or dysfunction in striatal dopamine receptor population. In summary, the homozygote tgHD showed a robust learning and memory impairment prior to any clear motor deficits, or striatal dysfunction. However, the data were not conclusive regarding how the memory systems were compromised and the precise nature and underlying mechanism of the cognitive deficit in the tgHD model requires further investigation.

Kirch RD; Meyer PT; Geisler S; Braun F; Gehrig S; Langen KJ; von Hörsten S; Nikkhah G; Cassel JC; Döbrössy MD

2013-02-01

147

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 (p47(phox) 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. PMID:18653711

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

2008-07-24

148

Renin inhibition attenuates insulin resistance, oxidative stress, and pancreatic remodeling in the transgenic Ren2 rat.  

UK PubMed Central (United Kingdom)

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 (p47(phox) 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 J; Whaley-Connell A; Hayden MR; DeMarco VG; Schneider R; Sowers SD; Karuparthi P; Ferrario CM; Sowers JR

2008-11-01

149

Twenty-one proteins up-regulated in human H-ras oncogene transgenic rat pancreas cancers are up-regulated in human pancreas cancer.  

UK PubMed Central (United Kingdom)

OBJECTIVES: We have established rat models of pancreatic ductal adenocarcinoma (PDAC) in which expression of a human H-ras(G12V) or K-ras(G12V) oncogene regulated by the Cre/lox system drives pancreatic carcinogenesis. Pancreatic ductal adenocarcinoma which develops in H-ras(G12V) and K-ras(G12V) transgenic rats is cytogenetically and histopathologically similar to human PDAC. The present study was designed to determine the feasibility of using the commercially available H-ras(G12V) transgenic rat to find diagnostic protein biomarkers for human pancreatic cancer. METHODS: For an animal model to be useful for searching for protein biomarkers for a disease, it is essential that proteins that are up-regulated in the model are also up-regulated in humans. We used liquid chromatography-tandem mass spectrometry (LC-MS/MS) to compare H-ras(G12V) transgenic rat PDAC with surrounding normal pancreas tissue. RESULTS: We identified 30 up-regulated proteins in the H-ras(G12V) transgenic rat PDAC lesions; importantly, 21 human homologs of these 30 rat proteins are up-regulated in human pancreatic cancer patients. CONCLUSIONS: These results indicate that numerous proteins that are up-regulated in H-ras(G12V) transgenic rat PDAC are also up-regulated in human pancreatic cancer; therefore, this rat model can be used to search for diagnostic biomarkers for this disease.

Yabushita S; Fukamachi K; Kikuchi F; Ozaki M; Miyata K; Sukata T; Deguchi Y; Tanaka H; Kakehashi A; Kawamura S; Uwagawa S; Wanibuchi H; Suzui M; Alexander DB; Tsuda H

2013-08-01

150

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

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

E Ferreira; E Potier; P Vaudin; K Oudina; M Bensidhoum; D Logeart-Avramoglou; LM Mir; H Petite

2012-01-01

151

Inhibitory Peptide of Mitochondrial ?-Calpain Protects against Photoreceptor Degeneration in Rhodopsin Transgenic S334ter and P23H Rats  

Science.gov (United States)

Mitochondrial ?-calpain and apoptosis-inducing factor (AIF)-dependent photoreceptor cell death has been seen in several rat and mouse models of retinitis pigmentosa (RP). Previously, we demonstrated that the specific peptide inhibitor of mitochondrial ?-calpain, Tat-µCL, protected against retinal degeneration following intravitreal injection or topical eye-drop application in Mertk gene-mutated Royal College of Surgeons rats, one of the animal models of RP. Because of the high rate of rhodopsin mutations in RP patients, the present study was performed to confirm the protective effects of Tat-µCL against retinal degeneration in rhodopsin transgenic S334ter and P23H rats. We examined the effects of intravitreal injection or topical application of the peptide on retinal degeneration in S334ter and P23H rats by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay, electroretinogram (ERG), immunohistochemistry for AIF, and histological staining. In S334ter rats, we found that intravitreal injection or topical application of the peptide prevented photoreceptor cell death from postnatal (PN) 15 to 18 days, the time of early-stage retinal degeneration. Topical application of the peptide also delayed attenuation of ERG responses from PN 28 to 56 days. In P23H rats, topical application of the peptide protected against photoreceptor cell death and nuclear translocation of AIF on PN 30, 40, and 50 days, as the primary stages of degeneration. We observed that topical application of the peptide inhibited the thinning of the outer nuclear layer and delayed ERG attenuations from PN 30 to 90 days. Our results demonstrate that the mitochondrial ?-calpain and AIF pathway is involved in early-stage retinal degeneration in rhodopsin transgenic S334ter and P23H rats, and inhibition of this pathway shows curative potential for rhodopsin mutation-caused RP.

Ozaki, Taku; Ishiguro, Sei-ichi; Hirano, Satoshi; Baba, Ayaka; Yamashita, Tetsuro; Tomita, Hiroshi; Nakazawa, Mitsuru

2013-01-01

152

Peritonitis activates transcription of the human prolactin locus in myeloid cells in a humanized transgenic rat model.  

UK PubMed Central (United Kingdom)

Prolactin (PRL) is mainly expressed in the pituitary in rodents, whereas in humans, expression is observed in many extrapituitary sites, including lymphocytes. Due to the lack of adequate experimental models, the function of locally produced PRL in the immune system is largely unknown. Using transgenic rats that express luciferase under the control of extensive human PRL regulatory regions, we characterized immune cell responses to thioglycollate (TG)-induced peritonitis. Resident populations of myeloid cells in the peritoneal cavity of untreated rats expressed barely detectable levels of luciferase. In contrast, during TG-induced peritonitis, cell-specific expression in both neutrophils and monocytes/macrophages in peritoneal exudates increased dramatically. Elevated luciferase expression was also detectable in peripheral blood and bone marrow CD11b(+) cells. Ex vivo stimulation of primary myeloid cells showed activation of the human extrapituitary promoter by TNF-?, lipopolysaccharide, or TG. These findings were confirmed in human peripheral blood monocytes, showing that the transgenic rat provided a faithful model for the human gene. Thus, the resolution of an inflammatory response is associated with dramatic activation of the PRL gene promoter in the myeloid lineage.

Semprini S; McNamara AV; Awais R; Featherstone K; Harper CV; McNeilly JR; Patist A; Rossi AG; Dransfield I; McNeilly AS; Davis JR; White MR; Mullins JJ

2012-06-01

153

Peritonitis activates transcription of the human prolactin locus in myeloid cells in a humanized transgenic rat model.  

Science.gov (United States)

Prolactin (PRL) is mainly expressed in the pituitary in rodents, whereas in humans, expression is observed in many extrapituitary sites, including lymphocytes. Due to the lack of adequate experimental models, the function of locally produced PRL in the immune system is largely unknown. Using transgenic rats that express luciferase under the control of extensive human PRL regulatory regions, we characterized immune cell responses to thioglycollate (TG)-induced peritonitis. Resident populations of myeloid cells in the peritoneal cavity of untreated rats expressed barely detectable levels of luciferase. In contrast, during TG-induced peritonitis, cell-specific expression in both neutrophils and monocytes/macrophages in peritoneal exudates increased dramatically. Elevated luciferase expression was also detectable in peripheral blood and bone marrow CD11b(+) cells. Ex vivo stimulation of primary myeloid cells showed activation of the human extrapituitary promoter by TNF-?, lipopolysaccharide, or TG. These findings were confirmed in human peripheral blood monocytes, showing that the transgenic rat provided a faithful model for the human gene. Thus, the resolution of an inflammatory response is associated with dramatic activation of the PRL gene promoter in the myeloid lineage. PMID:22495675

Semprini, Sabrina; McNamara, Anne V; Awais, Raheela; Featherstone, Karen; Harper, Claire V; McNeilly, Judith R; Patist, Amanda; Rossi, Adriano G; Dransfield, Ian; McNeilly, Alan S; Davis, Julian R E; White, Michael R H; Mullins, John J

2012-04-11

154

Activating the Nrf2-mediated antioxidant response element restores barrier function in the alveolar epithelium of HIV-1 transgenic rats.  

UK PubMed Central (United Kingdom)

The master transcription factor nuclear factor (erythroid-derived 2)-like 2 (Nrf2) regulates the expression of antioxidant and phase II-metabolizing enzymes by activating the antioxidant response element (ARE) and thereby protects cells and tissues from oxidative stress. Pulmonary complications remain the leading cause of death in human immunodeficiency virus (HIV)-1-infected individuals, who display systemic oxidative stress and glutathione deficiency that can be modeled in transgenic rats where HIV-1-related viral proteins decrease glutathione levels and cause epithelial barrier dysfunction within the alveolar space by as yet unknown mechanisms. We hypothesized that HIV-1-related proteins inhibit Nrf2-mediated antioxidant defenses and thereby disrupt the normally tight alveolar epithelial barrier. Nrf2 RNA silencing dampened Nrf2/ARE activity, decreased the expression of the tight junction proteins zonula occludens-1, occludin, and claudin-18, increased paracellular permeability of alveolar epithelial monolayers derived from wild-type rats, and therefore reproduced the effects of HIV-1 transgene expression on the epithelial barrier that we had previously described. In contrast, upregulating Nrf2 activity, either by plasmid-mediated overexpression or treatment with the Nrf2 activator sulforaphane, increased the expression of ARE-dependent antioxidants, including NAD(P)H dehydrogenase, quinone 1 and glutathione, improved the expression of tight junction proteins, and restored the ability to form tight barriers in alveolar epithelial cells from HIV-1 transgenic rats. Taken together, these new findings argue that HIV-1-related proteins downregulate Nrf2 expression and/or activity within the alveolar epithelium, which in turn impairs antioxidant defenses and barrier function, thereby rendering the lung susceptible to oxidative stress and injury. Furthermore, this study suggests that activating the Nrf2/ARE pathway with the dietary supplement sulforaphane could augment antioxidant defenses and lung health in HIV-1-infected individuals.

Fan X; Staitieh BS; Jensen JS; Mould KJ; Greenberg JA; Joshi PC; Koval M; Guidot DM

2013-08-01

155

A novel transgenic rat model for spinocerebellar ataxia type 17 recapitulates neuropathological changes and supplies in vivo imaging biomarkers.  

UK PubMed Central (United Kingdom)

Spinocerebellar ataxia 17 (SCA17) is an autosomal-dominant, late-onset neurodegenerative disorder caused by an expanded polyglutamine (polyQ) repeat in the TATA-box-binding protein (TBP). To further investigate this devastating disease, we sought to create a first transgenic rat model for SCA17 that carries a full human cDNA fragment of the TBP gene with 64 CAA/CAG repeats (TBPQ64). In line with previous observations in mouse models for SCA17, TBPQ64 rats show a severe neurological phenotype including ataxia, impairment of postural reflexes, and hyperactivity in early stages followed by reduced activity, loss of body weight, and early death. Neuropathologically, the severe phenotype of SCA17 rats was associated with neuronal loss, particularly in the cerebellum. Degeneration of Purkinje, basket, and stellate cells, changes in the morphology of the dendrites, nuclear TBP-positive immunoreactivity, and axonal torpedos were readily found by light and electron microscopy. While some of these changes are well recapitulated in existing mouse models for SCA17, we provide evidence that some crucial characteristics of SCA17 are better mirrored in TBPQ64 rats. Thus, this SCA17 model represents a valuable tool to pursue experimentation and therapeutic approaches that may be difficult or impossible to perform with SCA17 transgenic mice. We show for the first time positron emission tomography (PET) and diffusion tensor imaging (DTI) data of a SCA animal model that replicate recent PET studies in human SCA17 patients. Our results also confirm that DTI are potentially useful correlates of neuropathological changes in TBPQ64 rats and raise hope that DTI imaging could provide a biomarker for SCA17 patients.

Kelp A; Koeppen AH; Petrasch-Parwez E; Calaminus C; Bauer C; Portal E; Yu-Taeger L; Pichler B; Bauer P; Riess O; Nguyen HP

2013-05-01

156

Response to metal stress of Nicotiana langsdorffii plants wild-type and transgenic for the rat glucocorticoid receptor gene.  

UK PubMed Central (United Kingdom)

Recently our findings have shown that the integration of the gene coding for the rat gluco-corticoid receptor (GR receptor) in Nicotiana langsdorffii plants induced morphophysiological effects in transgenic plants through the modification of their hormonal pattern. Phytohormones play a key role in plant responses to many different biotic and abiotic stresses since a modified hormonal profile up-regulates the activation of secondary metabolites involved in the response to stress. In this work transgenic GR plants and isogenic wild type genotypes were exposed to metal stress by treating them with 30ppm cadmium(II) or 50ppm chromium(VI). Hormonal patterns along with changes in key response related metabolites were then monitored and compared. Heavy metal up-take was found to be lower in the GR plants. The transgenic plants exhibited higher values of S-abscisic acid (S-ABA) and 3-indole acetic acid (IAA), salicylic acid and total polyphenols, chlorogenic acid and antiradical activity, compared to the untransformed wild type plants. Both Cd and Cr treatments led to an increase in hormone concentrations and secondary metabolites only in wild type plants. Analysis of the results suggests that the stress responses due to changes in the plant's hormonal system may derive from the interaction between the GR receptor and phytosteroids, which are known to play a key role in plant physiology and development.

Fuoco R; Bogani P; Capodaglio G; Del Bubba M; Abollino O; Giannarelli S; Spiriti MM; Muscatello B; Doumett S; Turetta C; Zangrando R; Zelano V; Buiatti M

2013-05-01

157

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.

Joshi Pratibha C; Guidot David M

2011-01-01

158

Effects of 90-day feeding of transgenic Bt rice TT51 on the reproductive system in male rats.  

UK PubMed Central (United Kingdom)

Rice is a staple food crop; however, the threat of pests leads to a serious decline in its output and quality. The CryAb/CryAc gene, encodes a synthetic fusion Bacillus thuringiensis (Bt) crystal protein, was introduced into rice MingHui63 to produce insect-resistant rice TT51. This study was undertaken to investigate potential unintended effects of TT51 on the reproductive system in male rats. Male rats were treated with diets containing 60% of either TT51 or MingHui63 by weight, nutritionally balanced to an AIN93G diet, for 90days. An additional negative control group of rats were fed with a rice-based AIN93G diet. Body weights, food intake, hematology, serum chemistry, serum hormone levels, sperm parameters and relative organ/body weights were measured, and gross as well as microscopic pathology were examined. No diet-related significant differences in the values of response variables were observed between rats that were fed with diet containing transgenic TT51, MingHui63 and the control in this 90-day feeding study. In addition, necropsy and histopathology examination indicated no treatment-related changes. The results from the present study indicated that TT51 does not appear to exert any effect on the reproductive system in male rats compared with MingHui63 or the control.

Wang EH; Yu Z; Hu J; Xu HB

2013-09-01

159

Synthetic microRNA-mediated downregulation of Nogo-A in transgenic rats reveals its role as regulator of synaptic plasticity and cognitive function.  

UK PubMed Central (United Kingdom)

We have generated a transgenic rat model using RNAi and used it to study the role of the membrane protein Nogo-A in synaptic plasticity and cognition. The membrane protein Nogo-A is expressed in CNS oligodendrocytes and subpopulations of neurons, and it is known to suppress neurite growth and regeneration. The constitutively expressed polymerase II-driven transgene was composed of a microRNA-targeting Nogo-A placed into an intron preceding the coding sequence for EGFP, thus quantitatively labeling cells according to intracellular microRNA expression. The transgenic microRNA in vivo efficiently reduced the concentration of Nogo-A mRNA and protein preferentially in neurons. The resulting significant increase in long-term potentiation in both hippocampus and motor cortex indicates a repressor function of Nogo-A in synaptic plasticity. The transgenic rats exhibited prominent schizophrenia-like behavioral phenotypes, such as perseveration, disrupted prepulse inhibition, and strong withdrawal from social interactions. This fast and efficient microRNA-mediated knockdown provides a way to silence gene expression in vivo in transgenic rats and shows a role of Nogo-A in regulating higher cognitive brain functions.

Tews B; Schönig K; Arzt ME; Clementi S; Rioult-Pedotti MS; Zemmar A; Berger SM; Schneider M; Enkel T; Weinmann O; Kasper H; Schwab ME; Bartsch D

2013-04-01

160

Overexpression of HIF-1? transgene in the renal medulla attenuated salt sensitive hypertension in Dahl S rats.  

UK PubMed Central (United Kingdom)

Hypoxia inducible factor (HIF)-1?-mediated gene activation in the renal medulla in response to high salt intake plays an important role in the control of salt sensitivity of blood pressure. High salt-induced activation of HIF-1? in the renal medulla is blunted in Dahl S rats. The present study determined whether the impairment of the renal medullary HIF-1? pathway was responsible for salt sensitive hypertension in Dahl S rats. Renal medullary HIF-1? levels were induced by either transfection of HIF-1? expression plasmid or chronic infusion of CoCl? into the renal medulla, which was accompanied by increased expressions of anti-hypertensive genes, cyclooxygenase-2 and heme oxygenase-1. Overexpression of HIF-1? transgenes in the renal medulla enhanced the pressure natriuresis, promoted the sodium excretion and reduced sodium retention after salt overload. As a result, hypertension induced by 2-week high salt was significantly attenuated in rats treated with HIF-1? plasmid or CoCl?. These results suggest that an abnormal HIF-1? in the renal medulla may represent a novel mechanism mediating salt-sensitive hypertension in Dahl S rats and that induction of HIF-1? levels in the renal medulla could be a therapeutic approach for the treatment of salt-sensitive hypertension.

Zhu Q; Wang Z; Xia M; Li PL; Zhang F; Li N

2012-06-01

 
 
 
 
161

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

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

Sulie L. Chang; Michael Vigorito

2006-01-01

162

Regional variations of antioxidant capacity and oxidative stress responses in HIV-1 transgenic rats with and without methamphetamine administration.  

UK PubMed Central (United Kingdom)

HIV infection and methamphetamine (Meth) abuse both may lead to oxidative stress. This study used HIV-1 transgenic (HIV-1Tg) rats to investigate the independent and combined effects of HIV viral protein expression and low dose repeated Meth exposure on the glutathione (GSH)-centered antioxidant system and oxidative stress in the brain. Total GSH content, gene expression and/or enzymatic activities of glutamylcysteine synthetase (GCS), gamma-glutamyltransferase (GGT), glutathione reductase (GR), glutathione peroxidase (GPx), glutaredoxin (Glrx), and glutathione-s-transferase (GST) were measured. The protein expression of cystine transporter (xCT) and oxidative stress marker 4-hydroxynonenal (HNE) were also analyzed. Brain regions studied include thalamus, frontal and remainder cortex, striatum, cerebellum and hippocampus. HIV-1Tg rats and Meth exposure showed highly regional specific responses. In the F344 rats, the thalamus had the highest baseline GSH concentration and potentially higher GSH recycle rate. HIV-1Tg rats showed strong transcriptional responses to GSH depletion in the thalamus. Both HIV-1Tg and Meth resulted in decreased GR activity in thalamus, and decreased Glrx activity in frontal cortex. However, the increased GR and Glrx activities synergized with increased GSH concentration, which might have partially prevented Meth-induced oxidative stress in striatum. Interactive effects between Meth and HIV-1Tg were observed in thalamus on the activities of GCS and GGT, and in thalamus and frontal cortex on Glrx activity and xCT protein expression. Findings suggest that HIV viral protein and low dose repeated Meth exposure have separate and combined effects on the brain's antioxidant capacity and the oxidative stress response that are regional specific.

Pang X; Panee J; Liu X; Berry MJ; Chang SL; Chang L

2013-06-01

163

Incisor enamel formation is impaired in transgenic rats overexpressing the type III NaPi transporter Slc20a1.  

UK PubMed Central (United Kingdom)

Inorganic phosphate (Pi) is required in many biological processes, including signaling cascades, skeletal development, tooth mineralization, and nucleic acid synthesis. Recently, we showed that Pi transport in osteoblasts, mediated by Slc20a1, a member of the type III sodium-dependent phosphate transporter family, is indispensable for osteoid mineralization in rapidly growing rat bone. In addition, we found that bone mineral density decreased slightly with dysfunction of Pi homeostasis in aged transgenic rats overexpressing mouse Slc20a1 (Slc20a1-Tg). Bone and tooth share certain common molecular features, and thus, we focused on tooth development in Slc20a1-Tg mandibular incisors in order to determine the role of Slc20a1 in tooth mineralization. Around the time of weaning, there were no significant differences in serologic parameters between wild-type and Slc20a1-Tg rats. However, histological analysis showed that Slc20a1-Tg ameloblasts formed clusters in the papillary layer during the maturation stage as early as 4 weeks of age. These pathologies became more severe with age and included the formation of cyst-like or multilayer ameloblast structures, accompanied by a chalky white appearance with abnormal attrition and fracture. Hyperphosphatemia was also observed in aging Slc20a1-Tg rats. Micro-computed tomography and electron probe microanalysis revealed impairments in enamel, such as delayed mineralization and hypomineralization. Our results suggest that enamel formation is sensitive to imbalances in Pit1-mediated cellular function as seen in bone, although these processes are under the control of systemic Pi homeostasis.

Yoshioka H; Yoshiko Y; Minamizaki T; Suzuki S; Koma Y; Nobukiyo A; Sotomaru Y; Suzuki A; Itoh M; Maeda N

2011-09-01

164

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

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This study focuses on the effects of retinal degeneration on the circadian patterns of P23H rats, as well as on the effect of exogenous melatonin administration. To this end, the body temperature of P23H and Sprague–Dawley rats was continuously monitored and their retinas examined at different stage...

Lax Zapata, Pedro; Baño Otalora, Beatriz; Esquiva Sobrino, Gema; Rol de Lama, María de los Ángeles; Madrid Pérez, Juan Antonio

165

Alterations in 5-HT2A receptor signaling in male and female transgenic rats over-expressing either Gq or RGS-insensitive Gq protein.  

Science.gov (United States)

Serotonin 2A (5-HT2A) receptors are coupled to Galphaq and Galpha11 proteins to activate phospholipase C (PLC). Regulators of G-protein signaling proteins (RGS) modulate G-protein signaling by accelerating the intrinsic GTPase activity of Galphaq and Galpha11. This study investigated the effects of over-expression of wild-type Galphaq proteins (Gq-Tg) and over-expression of RGS-insensitive Galphaq proteins (G188S, RGSi-Tg) on 5-HT2A receptor mediated signaling in transgenic rats. Over-expression of wild-type Galphaq and RGS insensitive mutant Galphaq did not produce significant alterations in the levels of Galpha11, RGS2, RGS4, RGS7, RGS16 or 5-HT2A proteins. RGSi-Tg rats had higher oxytocin and corticosterone responses to (-)DOI, a 5-HT2A/2C receptor agonist, compared to Gq-Tg rats. RGSi-Tg and Gq-Tg rats had higher ACTH responses to (-)DOI compared to control rats. Similarly, 5-HT-stimulated PLC activity in the frontal cortex was higher in RGSi-Tg and Gq-Tg rats compared to control rats. In contrast, GTPgammaS-stimulated PLC activity was higher in Gq-Tg rats but not in RGSi-Tg rats compared to control rats. There was a small but statistically significant increase in the affinity of [125I]-DOI labeled 5-HT2A receptors in RGSi-Tg rats and Gq-Tg rats compared to controls. There were no significant differences in Bmax and Kd of [3H] ketanserin labeled 5-HT2A receptors among the three groups. These data suggest that the effect of RGS proteins on 5-HT2A receptor signaling is cell type specific. In transgenic rats over-expressing Galphaq, endogenous RGS proteins have a negative effect on 5-HT2A receptor-mediated oxytocin release. In contrast, endogenous RGS protein had no impact on 5-HT2A receptor-mediated ACTH release in transgenic rats. PMID:16769091

Shi, Ju; Damjanoska, Katerina J; Zemaitaitis, Bozena; Garcia, Francisca; Carrasco, Gonzalo; Sullivan, Nicole R; She, Yijin; Young, Kathleen H; Battaglia, George; Van De kar, Louis D; Howland, David S; Muma, Nancy A

2006-09-01

166

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

International Nuclear Information System (INIS)

[en] 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

167

Meniscal repair using bone marrow-derived mesenchymal stem cells: experimental study using green fluorescent protein transgenic rats.  

UK PubMed Central (United Kingdom)

Meniscal tears in the avascular zone have very limited potential to heal because of a poor blood supply. Although there have been many attempts to promote the healing potential of the torn meniscus, no established treatments have achieved sufficient meniscal healing. In this study, we evaluated the efficacy of mesenchymal stem cell transplantation as a cell source to promote meniscal healing, using cells from the green fluorescent protein (GFP) transgenic rat and organ culture model. Mesenchymal stem cells from bone marrow were isolated and expanded in monolayer culture. They were embedded in fibrin glue and were transplanted into the meniscal defects of Sprague-Dawley rats. In the control groups, the defects remained untreated, or only fibrin glue without cells was transplanted. The GFP-positive cells enabled us to detect the transplanted cells from recipient cells easily. As a result, transplanted mesenchymal stem cells could survive and proliferate in the meniscal defects in the organ culture model. They also could produce an abundant extracellular matrix stained by toluidine blue around the cells which contributed to meniscal healing in the avascular status. We could detect transplanted GFP cells under a fluorescent microscope until 8 weeks after transplantation. In a clinical situation, mesenchymal stem cell transplantation is a promising new clinical strategy for the treatment of meniscal tears in the avascular zone.

Izuta Y; Ochi M; Adachi N; Deie M; Yamasaki T; Shinomiya R

2005-06-01

168

Meniscal repair using bone marrow-derived mesenchymal stem cells: experimental study using green fluorescent protein transgenic rats.  

Science.gov (United States)

Meniscal tears in the avascular zone have very limited potential to heal because of a poor blood supply. Although there have been many attempts to promote the healing potential of the torn meniscus, no established treatments have achieved sufficient meniscal healing. In this study, we evaluated the efficacy of mesenchymal stem cell transplantation as a cell source to promote meniscal healing, using cells from the green fluorescent protein (GFP) transgenic rat and organ culture model. Mesenchymal stem cells from bone marrow were isolated and expanded in monolayer culture. They were embedded in fibrin glue and were transplanted into the meniscal defects of Sprague-Dawley rats. In the control groups, the defects remained untreated, or only fibrin glue without cells was transplanted. The GFP-positive cells enabled us to detect the transplanted cells from recipient cells easily. As a result, transplanted mesenchymal stem cells could survive and proliferate in the meniscal defects in the organ culture model. They also could produce an abundant extracellular matrix stained by toluidine blue around the cells which contributed to meniscal healing in the avascular status. We could detect transplanted GFP cells under a fluorescent microscope until 8 weeks after transplantation. In a clinical situation, mesenchymal stem cell transplantation is a promising new clinical strategy for the treatment of meniscal tears in the avascular zone. PMID:15911296

Izuta, Yasunori; Ochi, Mitsuo; Adachi, Nobuo; Deie, Masataka; Yamasaki, Takuma; Shinomiya, Rikuo

2005-01-07

169

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

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 con...

Charreau, B.; Ménoret, S.; Tesson, L.; Azimzadeh, A.; Audet, M.; Wolf, P.; Marquet, R.; Verbakel, C.; Ijzermans, J.

170

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

2013-05-21

171

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

UK PubMed Central (United Kingdom)

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

Lax P; Otalora BB; Esquiva G; Rol Mde L; Madrid JA; Cuenca N

2011-03-01

172

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

Science.gov (United States)

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

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

2010-11-09

173

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

Science.gov (United States)

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; P<0.05). The M/L (media-to-lumen) ratio assessed on pressurized preparations was equally reduced in aliskiren-treated and ramipril-treated rats compared with controls (6.3±0.5 and 6.4±0.2% compared with 9.8±0.4% respectively; P<0.05). Endothelium-dependent and -independent relaxations were similar among the groups. L-NAME (N(G)-nitro-L-arginine methyl ester) significantly reduced acetylcholine-induced dilation in drug-treated dTGR. This effect was significantly more prominent in aliskiren-treated rats. eNOS (endothelial NO synthase) expression showed a 2-fold increase only in aliskiren-treated dTGR as compared with controls (P<0.01) and ramipril-treated dTGR (P<0.05). Plasma nitrite, as an index of NO production, was significantly increased in dTGR treated with either aliskiren or ramipril compared with controls. Only aliskiren induced a 2-fold increase in plasma nitrite, which was significantly greater than that induced by ramipril (P<0.05). gp91(phox) expression and ROS (reactive oxygen species) production in aorta were significantly and similarly reduced by both drugs. In conclusion, equieffective hypotensive doses of aliskiren or ramipril 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

174

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

175

An efficient expression of Human Growth Hormone (hGH) in the milk of transgenic mice using rat {beta}-casein/hGH fusion genes  

Energy Technology Data Exchange (ETDEWEB)

In order to produce human growth hormone (hGH) in the milk of transgenic mice, two expression vectors for hGH differing in their 3{prime} flanking sequences were constructed by placing the genomic sequences of hGH gene under the control of the rat {beta}-casein gene promotor. The 3{prime} flanking sequences of the expression constructs were derived from either the hGH gene (pBCN1GH) or the rat {beta}-casein gene (pBCN2GH). Transgenic lines bearing pBCN1GH expressed hGH more efficiently than those bearing pBCN2GH in the milk (19-5500 {mu}g/mL vs 0.7-2 {mu}g/mL). In particular, one of the BCN1GH lines expressed hGH as much as 5500 {plus_minus} 620 {mu}g/mL. Northern blot analysis showed that the transgene expression was specifically confined to the mammary gland and developmentally regulated like the endogeneous mouse {beta}-casein gene in the mammary gland. However, a low level of nonmammary expression was also detected with more sensitive assay methods. In conclusion, the rat {beta}-casein/hGH fusion gene could direct an efficient production of hGH in a highly tissue- and stage-specific manner in the transgenic mice and the 3{prime} flanking sequences of hGH gene had an important role for the efficient expression. 27 refs., 5 figs., 2 tabs.

Lee, Chul-Sang; Yu, Dae-Yeul; Lee, Kyung-Kwang [Korea Research Institute of Bioscience and Biotechnology, Taejon (Korea, Republic of)] [and others

1996-03-01

176

Single intra-articular injection of adeno-associated virus results in stable and controllable in vivo transgene expression in normal rat knees.  

UK PubMed Central (United Kingdom)

OBJECTIVE: To test the hypothesis that in vivo transgene expression mediated by single intra-articular injection of adeno-associated virus serotype 2 (AAV2) persists within intra-articular tissues 1 year post-injection and can be externally controlled using an AAV2-based tetracycline-inducible gene regulation system containing the tetracycline response element (TRE) promoter. METHODS: Sprague Dawley rats received intra-articular injections of AAV2-cytomegalovirus (CMV)-enhanced green fluorescent protein (GFP) and AAV2-CMV-luciferase (Luc) into their right and left knees, respectively. Luciferase expression was evaluated over 1 year using bioluminescence imaging. After sacrifice, tissues were analyzed for GFP+ cells by fluorescent microscopy. To study external control of intra-articular AAV-transgene expression, another set of rats was co-injected with AAV2-TRE-Luc and AAV2-CMV-reverse-tetracycline-controlled transactivator (rtTA) into the right knees, and AAV2-CMV-Luc and AAV2-CMV-rtTA into the left knees. Rats received oral doxycycline (Dox), an analog of tetracycline, for 7 days. Luciferase expression was assessed by bioluminescence imaging. RESULTS: Luciferase expression was localized to the injected joint and persisted throughout the 1-year study period. Abundant GFP+ cells were observed within intra-articular soft tissues. Transgene expression in AAV2-TRE-Luc injected joints was upregulated by oral administration of Dox, and downregulated following its removal, at 14 days and 13 months post-AAV injection. CONCLUSIONS: This longitudinal in vivo study shows that sustained and stable AAV-mediated intra-articular transgene expression can be achieved through a single intra-articular injection and can be controlled using a tetracycline-controlled inducible AAV system in a normal rat knee model. Highly regulatable long-term intra-articular transgene expression is of potential clinical utility for development of treatment strategies for chronic intra-articular disease processes such as inflammatory and degenerative arthritis.

Payne KA; Lee HH; Haleem AM; Martins C; Yuan Z; Qiao C; Xiao X; Chu CR

2011-08-01

177

Sustained transgene expression in rat kidney with naked plasmid DNA and PCR-amplified DNA fragments.  

UK PubMed Central (United Kingdom)

Recently, we developed a kidney-targeted gene transfer technique, in which naked DNA was injected into the renal vein while the renal vein and artery were clamped. Kidney-targeted DNA transfer with only the renal vein clamped is an important modification that may permit less invasive catheter-based gene transfer in future clinical applications. The preparation of PCR-amplified DNA fragments is less time-consuming than that of naked plasmid DNA. We examined rat erythropoietin (Epo) plasmid, pCAGGS-Epo, or PCR-amplified DNA fragment, fCAGGS-Epo, transfer into the rat kidney with only the renal vein clamped. The Epo level peaked at week 3 and then was sustained for 24 weeks, which resulted in significant erythropoiesis. This modified technique, allowing long-term expression of both PCR-amplified DNA fragments and naked plasmid DNA, could potentially be used for catheter-based gene transfer in humans, and could help determine the physiological functions of putative genes.

Maruyama H; Higuchi N; Kameda S; Nakamura G; Shimotori M; Iino N; Higuchi M; Neichi T; Yokoyama S; Kono T; Miyazaki J; Gejyo F

2005-03-01

178

Sustained transgene expression in rat kidney with naked plasmid DNA and PCR-amplified DNA fragments.  

Science.gov (United States)

Recently, we developed a kidney-targeted gene transfer technique, in which naked DNA was injected into the renal vein while the renal vein and artery were clamped. Kidney-targeted DNA transfer with only the renal vein clamped is an important modification that may permit less invasive catheter-based gene transfer in future clinical applications. The preparation of PCR-amplified DNA fragments is less time-consuming than that of naked plasmid DNA. We examined rat erythropoietin (Epo) plasmid, pCAGGS-Epo, or PCR-amplified DNA fragment, fCAGGS-Epo, transfer into the rat kidney with only the renal vein clamped. The Epo level peaked at week 3 and then was sustained for 24 weeks, which resulted in significant erythropoiesis. This modified technique, allowing long-term expression of both PCR-amplified DNA fragments and naked plasmid DNA, could potentially be used for catheter-based gene transfer in humans, and could help determine the physiological functions of putative genes. PMID:15809339

Maruyama, Hiroki; Higuchi, Noboru; Kameda, Shigemi; Nakamura, Gen; Shimotori, Masaaki; Iino, Noriaki; Higuchi, Masato; Neichi, Tomohiro; Yokoyama, Sadaaki; Kono, Toru; Miyazaki, Jun-ichi; Gejyo, Fumitake

2005-03-01

179

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

180

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

Directory of Open Access Journals (Sweden)

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.

Jeyaraj D Antony; Grossman Gail; Petrusz Peter

2003-01-01

 
 
 
 
181

Renal nitric oxide synthase and antioxidant preservation in cyp1a1-ren-2 transgenic rats with inducible malignant hypertension.  

UK PubMed Central (United Kingdom)

BACKGROUND: Dietary administration of 0.30% indole-3-carbinol (I3C) to Cyp1a1-Ren2 transgenic rats (TGRs) generates angiotensin II (ANG II)-dependent malignant hypertension (HTN) and increased renal vascular resistance. However, TGRs with HTN maintain a normal or slightly reduced glomerular filtration rate. We tested the hypothesis that maintenance of renal function in hypertensive Cyp1a1-Ren2 TGRs is due to preservation of the intrarenal nitric oxide (NO) and antioxidant systems. METHODS: Kidney cortex, kidney medulla, aortic endothelial (e) and neuronal (n) nitric oxide synthase (NOS), superoxide dismutases (SODs), and p22phox (nicotinamide adenine dinucleotide phosphate-oxidase subunit) protein abundances were measured along with kidney cortex total antioxidant capacity (TAC) and NOx. TGRs were fed a normal diet that contained 0.3% I3C or 0.3% I3C + candesartan (AT1 receptor antagonist; 25mg/L in drinking water) (n = 5-6 per group) for 10 days. RESULTS: Blood pressure increased and body weight decreased in I3C-induced TGRs, while candesartan blunted these responses. Abundances of NOS, SOD, and p22phox as well as TAC were maintained in the kidney cortex of I3C-induced TGRs with and without candesartan, while kidney cortex NOx production increased in both groups. Kidney medulla eNOS and extracellular (EC) SOD decreased and nNOS were unchanged in both groups of I3C-induced TGRs. In addition, a compensatory increase occurred in kidney medulla Mn SOD in I3C-induced TGRs + candesartan. Aortic eNOS and nNOS? fell and p22phox and Mn SOD increased in hypertensive I3C-induced TGRs; all changes were reversed with candesartan. CONCLUSIONS: The preservation of renal cortical NO and antioxidant capacity is associated with preserved renal function in Cyp1a1-Ren2 TGRs with ANG II-dependent malignant HTN.

Cunningham MW Jr; Sasser JM; West CA; Milani CJ; Baylis C; Mitchell KD

2013-10-01

182

Renal Nitric Oxide Synthase and Antioxidant Preservation in Cyp1a1-Ren-2 Transgenic Rats With Inducible Malignant Hypertension.  

UK PubMed Central (United Kingdom)

BACKGROUND: Dietary administration of 0.30% indole-3-carbinol (I3C) to Cyp1a1-Ren2 transgenic rats (TGRs) generates angiotensin II (ANG II)-dependent malignant hypertension (HTN) and increased renal vascular resistance. However, TGRs with HTN maintain a normal or slightly reduced glomerular filtration rate. We tested the hypothesis that maintenance of renal function in hypertensive Cyp1a1-Ren2 TGRs is due to preservation of the intrarenal nitric oxide (NO) and antioxidant systems. METHODS: Kidney cortex, kidney medulla, aortic endothelial (e) and neuronal (n) nitric oxide synthase (NOS), superoxide dismutases (SODs), and p22phox (nicotinamide adenine dinucleotide phosphate-oxidase subunit) protein abundances were measured along with kidney cortex total antioxidant capacity (TAC) and NOx. TGRs were fed a normal diet that contained 0.3% I3C or 0.3% I3C + candesartan (AT1 receptor antagonist; 25mg/L in drinking water) (n = 5-6 per group) for 10 days. RESULTS: Blood pressure increased and body weight decreased in I3C-induced TGRs, while candesartan blunted these responses. Abundances of NOS, SOD, and p22phox as well as TAC were maintained in the kidney cortex of I3C-induced TGRs with and without candesartan, while kidney cortex NOx production increased in both groups. Kidney medulla eNOS and extracellular (EC) SOD decreased and nNOS were unchanged in both groups of I3C-induced TGRs. In addition, a compensatory increase occurred in kidney medulla Mn SOD in I3C-induced TGRs + candesartan. Aortic eNOS and nNOS? fell and p22phox and Mn SOD increased in hypertensive I3C-induced TGRs; all changes were reversed with candesartan. CONCLUSIONS: The preservation of renal cortical NO and antioxidant capacity is associated with preserved renal function in Cyp1a1-Ren2 TGRs with ANG II-dependent malignant HTN.

Cunningham MW Jr; Sasser JM; West CA; Milani CJ; Baylis C; Mitchell KD

2013-06-01

183

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

UK PubMed Central (United Kingdom)

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.

Amer DA; Kretzschmar G; Müller N; Stanke N; Lindemann D; Vollmer G

2010-06-01

184

Antihypertensive action of soluble epoxide hydrolase inhibition in Ren-2 transgenic rats is mediated by suppression of the intrarenal renin-angiotensin system.  

UK PubMed Central (United Kingdom)

The aim of the present study was to evaluate the hypothesis that the antihypertensive effects of inhibition of soluble epoxide hydrolase (sEH) are mediated by increased intrarenal availability of epoxyeicosatrienoic acids (EETs), with consequent improvement in renal haemodynamic autoregulatory efficiency and the pressure-natriuresis relationship. Ren-2 transgenic rats (TGR), a model of angiotensin (Ang) II-dependent hypertension, and normotensive transgene-negative Hannover Sprague-Dawley (HanSD) rats were treated with the sEH inhibitor cis-4-(4-(3-adamantan-1-yl-ureido)cyclohexyloxy)benzoic acid (c-AUCB; 26 mg/L) for 48 h. Then, the effects on blood pressure (BP), autoregulation of renal blood flow (RBF) and glomerular filtration rate (GFR), and on the pressure-natriuresis relationship in response to stepwise reductions in renal arterial pressure (RAP) were determined. Treatment with c-AUCB did not significantly change BP, renal autoregulation or pressure-natriuresis in normotensive HanSD rats. In contrast, c-AUCB treatment significantly reduced BP, increased intrarenal bioavailability of EETs and significantly suppressed AngII levels in TGR. However, treatment with c-AUCB did not significantly improve the autoregulatory efficiency of RBF and GFR in response to reductions of RAP and to restore the blunted pressure-natriuresis relationship in TGR. Together, the data indicate that the antihypertensive actions of sEH inhibition in TGR are predominantly mediated via significant suppression of intrarenal renin-angiotensin system activity.

Varcabova S; Huskova Z; Kramer HJ; Hwang SH; Hammock BD; Imig JD; Kitada K; Cervenka L

2013-04-01

185

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)

[en] 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

186

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

UK PubMed Central (United Kingdom)

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.

Cao S; He X; Xu W; Luo Y; Yuan Y; Liu P; Cao B; Shi H; Huang K

2012-03-01

187

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

2012-01-03

188

Transgenic TGR(mREN2)27 rats as a model for disturbed circadian organization at the level of the brain, the heart, and the kidneys.  

Science.gov (United States)

In transgenic hypertensive TGR(mREN2)27 rats (TGR) harboring the murine Ren-2 gene an inverse 24h blood pressure (BP) profile was described in relation to a normal pattern in heart rate (HR) and motility (MA), normotensive Sprague-Dawley rats (SDR) were used as controls. Transgenic rats as an animal model of human secondary hypertension (non-dipper) was studied in detail at different levels: (1) Radiotelemetry was applied to document gross circadian rhythms/rhythm disturbances in cardiovascular functions, MA and body temperature under normal LD conditions, under DD and after a light pulse. (2) Signal transduction of the overexpressed renin-angiotensin in TGR was studied by determation of AT1-receptors in kidney glomeruli together with kidney functions. (3) Expression of key processes involved in increased sympathetic regulation in TGR, mRNAs, the tyrosine-hydroxylase (TH) and norepinephrine (NE) reuptake1-carrier were determined. (4) In the SCN mRNA of c-fos and c-jun were determined under LD and after light pulse. (5) In primary cultures of pinealocytes the effects of adrenergic agonists and antagonists were evaluated on second messenger (cAMP, cGMP) accumulation and melatonin release. The results of these studies clearly demonstrate that the additional mouse renin genin in TGR greatly affected not only the renin-angiotensin-system and led--as expected--to an increased BP in this rat but also disturbed circadian rhythms from the BP pattern down to the level of hormones, processes of signal transduction, and expression of transcription factors and clock genes. In conclusion, the expression of a single additional gene is able to disturb the circadian system of an animal in a highly complex way. These findings are importance for chronobiologic as well as pharmacologic research. PMID:12916722

Lemmer, Björn; Witte, Klaus; Enzminger, Helene; Schiffer, Sabine; Hauptfleisch, Stefan

2003-07-01

189

The Wlds transgene reduces axon loss in a Charcot-Marie-Tooth disease 1A rat model and nicotinamide delays post-traumatic axonal degeneration.  

UK PubMed Central (United Kingdom)

Charcot-Marie-Tooth disease (CMT) is the most common inherited neuropathy and a duplication of the peripheral myelin protein of 22 kDa (PMP22) gene causes the most frequent subform CMT1A. Clinical impairments are determined by the amount of axonal loss. Axons of the spontaneous mouse mutant Wallerian degeneration slow (Wlds) show markedly reduced degeneration following various types of injuries. Protection is conferred by a chimeric Wlds gene encoding an N-terminal part of ubiquitination factor Ube4b and full length nicotinamide mononucleotide adenylyl transferase 1 (Nmnat1). Nmnat1 enzyme generates nicotinamide adenine dinucleotide (NAD) from nicotinamide mononucleotide. Here, in a Pmp22 transgenic animal model of Charcot-Marie-Tooth disease type 1A (CMT rat), the Wlds transgene reduced axonal loss and clinical impairments without altering demyelination. Furthermore, nicotinamide - substrate precursor of the Nmnat1 enzyme - transiently delayed posttraumatic axonal degeneration in an in vivo model of acute peripheral nerve injury, but to a lower extent than Wlds. In contrast, 8 weeks of nicotinamide treatment did not influence axonal loss or clinical manifestations in the CMT rat. Therefore, nicotinamide can partially substitute for the protective Wlds effect in acute traumatic, but not in chronic secondary axonal injury. Future studies are needed to develop axon protective therapy in CMT1A which may be combined with therapeutic strategies aimed at downregulation of toxic PMP22 overexpression.

Meyer zu Horste G; Miesbach TA; Muller JI; Fledrich R; Stassart RM; Kieseier BC; Coleman MP; Sereda MW

2011-04-01

190

The Wlds transgene reduces axon loss in a Charcot-Marie-Tooth disease 1A rat model and nicotinamide delays post-traumatic axonal degeneration.  

Science.gov (United States)

Charcot-Marie-Tooth disease (CMT) is the most common inherited neuropathy and a duplication of the peripheral myelin protein of 22 kDa (PMP22) gene causes the most frequent subform CMT1A. Clinical impairments are determined by the amount of axonal loss. Axons of the spontaneous mouse mutant Wallerian degeneration slow (Wlds) show markedly reduced degeneration following various types of injuries. Protection is conferred by a chimeric Wlds gene encoding an N-terminal part of ubiquitination factor Ube4b and full length nicotinamide mononucleotide adenylyl transferase 1 (Nmnat1). Nmnat1 enzyme generates nicotinamide adenine dinucleotide (NAD) from nicotinamide mononucleotide. Here, in a Pmp22 transgenic animal model of Charcot-Marie-Tooth disease type 1A (CMT rat), the Wlds transgene reduced axonal loss and clinical impairments without altering demyelination. Furthermore, nicotinamide - substrate precursor of the Nmnat1 enzyme - transiently delayed posttraumatic axonal degeneration in an in vivo model of acute peripheral nerve injury, but to a lower extent than Wlds. In contrast, 8 weeks of nicotinamide treatment did not influence axonal loss or clinical manifestations in the CMT rat. Therefore, nicotinamide can partially substitute for the protective Wlds effect in acute traumatic, but not in chronic secondary axonal injury. Future studies are needed to develop axon protective therapy in CMT1A which may be combined with therapeutic strategies aimed at downregulation of toxic PMP22 overexpression. PMID:21168501

Meyer zu Horste, Gerd; Miesbach, Timo A; Muller, Johanna I; Fledrich, Robert; Stassart, Ruth M; Kieseier, Bernd C; Coleman, Michael P; Sereda, Michael W

2010-12-16

191

Human cyclin T1 expression ameliorates a T-cell-specific transcriptional limitation for HIV in transgenic rats, but is not sufficient for a spreading infection of prototypic R5 HIV-1 strains ex vivo  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Cells derived from native rodents have limits at distinct steps of HIV replication. Rat primary CD4 T-cells, but not macrophages, display a profound transcriptional deficit that is ameliorated by transient trans-complementation with the human Tat-interacting protein Cyclin T1 (hCycT1). Results Here, we generated transgenic rats that selectively express hCycT1 in CD4 T-cells and macrophages. hCycT1 expression in rat T-cells boosted early HIV gene expression to levels approaching those in infected primary human T-cells. hCycT1 expression was necessary, but not sufficient, to enhance HIV transcription in T-cells from individual transgenic animals, indicating that endogenous cellular factors are critical co-regulators of HIV gene expression in rats. T-cells from hCD4/hCCR5/hCycT1-transgenic rats did not support productive infection of prototypic wild-type R5 HIV-1 strains ex vivo, suggesting one or more significant limitation in the late phase of the replication cycle in this primary rodent cell type. Remarkably, we identify a replication-competent HIV-1 GFP reporter strain (R7/3 YU-2 Env) that displays characteristics of a spreading, primarily cell-to-cell-mediated infection in primary T-cells from hCD4/hCCR5-transgenic rats. Moreover, the replication of this recombinant HIV-1 strain was significantly enhanced by hCycT1 transgenesis. The viral determinants of this so far unique replicative ability are currently unknown. Conclusion Thus, hCycT1 expression is beneficial to de novo HIV infection in a transgenic rat model, but additional genetic manipulations of the host or virus are required to achieve full permissivity.

Michel Nico; Goffinet Christine; Ganter Kerstin; Allespach Ina; KewalRamani Vineet N; Saifuddin Mohammed; Littman Dan R; Greene Warner C; Goldsmith Mark A; Keppler Oliver T

2009-01-01

192

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

193

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

Science.gov (United States)

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

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

2012-08-01

194

The nutritional evaluation of transgenic rice  

UK PubMed Central (United Kingdom)

Weaning Wistar rats were fed with food composed of transgenic rice (in which cowpea trypsin inhibitor gene was introduced) or nontransgenic rice for 28d. Their food had the same proportion of protein (10%) and most protein was the rice protein accordingly (a little casein added). The following nutritional indices were compared: blood chemisty, body weight and body length, organ index, PER, bone density. No differences were found in the nutritional indices between rats fed with transgenic rice and rats with nontransgenic rice. There is substantial equivalence of nutritive value of proteins between transgenic rice and nontransgenic rice. Cowpea trypsin inhibitor in transgenic rice has no bad impact on the absorptiono of other nutrients in diet. The transgenic rice is basically safe from nutritional aspect.

Chen Xiaoping; Zhuo Qin; Gu Lvzhen; Piao Jianhua; Yang Xiaoguang

2004-01-01

195

Transgenic tea  

Digital Repository Infrastructure Vision for European Research (DRIVER)

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 paramete...

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

196

Respiration and ROS production in brain and spinal cord mitochondria of transgenic rats with mutant G93a Cu/Zn-superoxide dismutase gene.  

UK PubMed Central (United Kingdom)

UNLABELLED: Mitochondrial dysfunction is involved in the pathogenesis of motor neuron degeneration in the G93A mutant transgenic (tgmSOD1) animal model of ALS. However, it is unknown whether mitochondriopathy is a primary or secondary event. We isolated brain (BM) and spinal cord (SCM) mitochondria from 2 month old presymptomatic tgmSOD1 rats and studied respiration and generation of reactive oxygen species (ROS) using a new metabolic paradigm (Panov et al., Am. J. Physiol., Regul. Integr. Comp. Physiol., 2011). The yields of BM and SCM from tgmSOD1 rats were 27% and 58% lower than normal rats (WT). The rates of the State 3 and State 3U respiration of tgBM and tgSCM were normal with glutamate+pyruvate+malate as substrates but were inhibited with pyruvate+malate in tgBM and glutamate+malate in tgSCM. In tgSCM the State 4 respiration with all substrates was significantly (1.5-2 fold) increased as compared with WT-SCM. Western blot analysis showed that tgSCM had lower contents of complexes III (-60%) and IV (-35%), and the presence of mutated SOD1 protein in both tgBM and tgSCM. With glutamate+pyruvate+malate or succinate+glutamate+pyruvate+malate as substrates, tgBM and tgSCM generated 5-7 fold more ROS than normal mitochondria, and tgSCM generated two times more ROS than tgBM. We show that the major damaging ROS species in tgmSOD1 animals is H(2)O(2). It is known that mutated SOD1, damaged by H(2)O(2), associates with mitochondria, and we suggest that this further increases production of H(2)O(2). We also show that the total tissue calcium content remained normal in the brain but was diminished by 26% in the spinal cord of presymptomatic tgmSOD1 rats. CONCLUSION: In tgSCM abnormally high rates of ROS generation, associated with reverse electron transport, result in accelerated mitochondriopathy, and the Ca(2+)-dependent excitotoxic death of motor neurons. Thus mitochondrial dysfunction is a key early element in pathogenesis of motor neuron degeneration in tgmSOD1 rats.

Panov A; Kubalik N; Zinchenko N; Hemendinger R; Dikalov S; Bonkovsky HL

2011-10-01

197

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.

El?bieta Musiej-Nowakowska; Barbara M?czy?ska-Rusiniak; Beata Ko?odziejczyk; Agnieszka Gazda

2011-01-01

198

Regressive and reactive changes in the connectivity patterns of rod and cone pathways of P23H transgenic rat retina  

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We have used the P23H line 1 homozygous albino rat to study how progressive photoreceptor degeneration affects rod and cone relay pathways. We examined P23H retinas at different stages of degeneration by confocal microscopy of immunostained sections and electroretinogram (ERG) recordings. By 21 day...

Cuenca Navarro, Nicolás; Pinilla Lozano, Isabel; Sauvé, Yves; Lu, B.; Wang, Shaomei; Lund, Raymond D.

199

Possible enhancing activity of diacylglycerol on 4-nitroquinoline 1-oxide induced carcinogenesis of the tongue in human c-Ha-ras proto-oncogene transgenic rats.  

Science.gov (United States)

1,2-diacylglycerol (1,2-DAG) is involved in cell proliferation as an activator of protein kinase C (PKC) and has been shown to stimulate growth of cancer cells, raising the possibility of a role in tumor promotion. Ingested DAG oil, containing 70% 1,3-DAG and 30% 1,2-DAG, is digested and considered to be safe as edible oil. However, DAG may directly contact with oral cavity mucosa in undigested form. The present study was conducted to examine the effects of DAG oil on carcinogenesis in c-Ha-ras proto-oncogene transgenic (Tg) rats administered 4-nitroquinoline 1-oxide (4NQO, 10 ppm) in their drinking water for 10 weeks for initiation of mainly upper digestive organs. DAG oil added in basal diet at 5.5%, 2.75%, 1.38% and 0% with total fat made up to 5.5% with triacylglycerol (TAG) was administered during the initiation and post-initiation period. The study was terminated at week 12 (Tg females) and 20 (Tg males, wild females and males). The fatty acid composition of DAG oil was similar to TAG (linoleic acid 46.6% and oleic acid 38.9%). In Tg male rats, DAG oil administration was associated with significant increase (P<0.05) in the incidence of squamous cell carcinomas (SCC) of the tongue (5.5% DAG, 43.8%; 2.75% DAG, 20%; 1.38% DAG, 14.3%; 0%, 12.3%) with the Cochran-Armitage trend test and also number of tumors in coefficients for linear contrast trend tests. Tongue SCC induction of wild males and all females was not significant. The present results suggest that DAG oil may have enhancing and/or promotion potential for tongue carcinogenesis in male Tg featuring elevated ras expression. PMID:17258375

Tsuda, Hiroyuki; Iigo, Masaaki; Takasuka, Nobuo; Ueda, Shinobu; Ohshima, Yutaka; Fukamachi, Katsumi; Shirai, Tomoyuki; Hirano, Sachiko; Matsuda, Eiji; Wakabayashi, Keiji

2006-12-20

200

Sperm parameters and epididymis function in transgenic rats overexpressing the Ca2+-binding protein regucalcin: a hidden role for Ca2+ in sperm maturation?  

Science.gov (United States)

Sperm undergo maturation acquiring progressive motility and the ability to fertilize oocytes through exposure to the components of the epididymal fluid (EF). Although the establishment of a calcium (Ca(2+)) gradient along the epididymis has been described, its direct effects on epididymal function remain poorly explored. Regucalcin (RGN) is a Ca(2+)-binding protein, regulating the activity of Ca(2+)-channels and Ca(2+)-ATPase, for which a role in male reproductive function has been suggested. This study aimed at comparing the morphology, assessed by histological analysis, and function of epididymis, by analysis of sperm parameters, antioxidant potential and Ca(2+) fluxes, between transgenic rats overexpressing RGN (Tg-RGN) and their wild-type littermates. Tg-RGN animals displayed an altered morphology of epididymis and lower sperm counts and motility. Tissue incubation with (45)Ca(2+) showed also that epididymis of Tg-RGN displayed a diminished rate of Ca(2+)-influx, indicating unbalanced Ca(2+) concentrations in the epididymal lumen. Sperm viability and the frequency of normal sperm, determined by the one-step eosin-nigrosin staining technique and the Diff-Quik staining method, respectively, were higher in Tg-RGN. Moreover, sperm of Tg-RGN rats showed a diminished incidence of tail defects. Western blot analysis demonstrated the presence of RGN in EF as well as its higher expression in the corpus region. The results presented herein demonstrated the importance of maintaining Ca(2+)-levels in the epididymal lumen and suggest a role for RGN in sperm maturation. Overall, a new insight into the molecular mechanisms driving epididymal sperm maturation was obtained, which could be relevant to development of better approaches in male infertility treatment and contraception. PMID:23615721

Correia, S; Oliveira, P F; Guerreiro, P M; Lopes, G; Alves, M G; Canário, A V M; Cavaco, J E; Socorro, Sílvia

2013-04-23

 
 
 
 
201

Transplantation of photoreceptor and total neural retina preserves cone function in P23H rhodopsin transgenic rat.  

UK PubMed Central (United Kingdom)

BACKGROUND: Transplantation as a therapeutic strategy for inherited retinal degeneration has been historically viewed to restore vision as a method by replacing the lost retinal cells and attempting to reconstruct the neural circuitry with stem cells, progenitor cells and mature neural retinal cells. METHODS AND FINDINGS: We present evidence for an alternative strategy aimed at preventing the secondary loss of cones, the most crucial photoreceptors for vision, by transplanting normal photoreceptors cells into the eye of the P23H rat, a model of dominant retinitis pigmentosa. We carried out transplantation of photoreceptors or total neural retina in 3-month-old P23H rats and evaluated the function and cell counts 6 months after surgery. In both groups, cone loss was significantly reduced (10%) in the transplanted eyes where the cone outer segments were found to be considerably longer. This morphological effect correlated with maintenance of the visual function of cones as scored by photopic ERG recording, but more precisely with an increase in the photopic b-wave amplitudes by 100% and 78% for photoreceptor transplantation and whole retinal transplantation respectively. CONCLUSIONS: We demonstrate here that the transplanted tissue prevents the loss of cone function, which is further translated into cone survival.

Yang Y; Mohand-Said S; Léveillard T; Fontaine V; Simonutti M; Sahel JA

2010-01-01

202

Endoplasmic reticulum stress and the unfolded protein response are linked to synergistic IFN-beta induction via X-box binding protein 1.  

Science.gov (United States)

Type I IFN are strongly induced upon engagement of certain pattern recognition receptors by microbial products, and play key roles in regulating innate and adaptive immunity. It has become apparent that the endoplasmic reticulum (ER) stress-induced unfolded protein response (UPR), in addition to restoring ER homeostasis, also influences the expression of certain inflammatory cytokines. However, the extent to which UPR signaling regulates type I IFN remains unclear. Here we show that cells undergoing a UPR respond to TLR4 and TLR3 ligands, and intracellular dsRNA, with log-fold greater IFN-beta induction. This synergy is not dependent on autocrine type I IFN signaling, but unexpectedly requires the UPR transcription factor X-box binding protein 1 (XBP-1). Synergistic IFN-beta induction also occurs in HLA-B27/human beta(2)m-transgenic rat macrophages exhibiting a UPR as a consequence of HLA-B27 up-regulation, where it correlates with activation of XBP-1 splicing. Together these findings indicate that the cellular response to endogenous 'danger' that disrupts ER homeostasis is coupled to IFN-beta induction by XBP-1, which has implications for the immune response and the pathogenesis of diseases involving the UPR. PMID:18412159

Smith, Judith A; Turner, Matthew J; DeLay, Monica L; Klenk, Erin I; Sowders, Dawn P; Colbert, Robert A

2008-05-01

203

Endoplasmic reticulum stress and the unfolded protein response are linked to synergistic IFN-beta induction via X-box binding protein 1.  

UK PubMed Central (United Kingdom)

Type I IFN are strongly induced upon engagement of certain pattern recognition receptors by microbial products, and play key roles in regulating innate and adaptive immunity. It has become apparent that the endoplasmic reticulum (ER) stress-induced unfolded protein response (UPR), in addition to restoring ER homeostasis, also influences the expression of certain inflammatory cytokines. However, the extent to which UPR signaling regulates type I IFN remains unclear. Here we show that cells undergoing a UPR respond to TLR4 and TLR3 ligands, and intracellular dsRNA, with log-fold greater IFN-beta induction. This synergy is not dependent on autocrine type I IFN signaling, but unexpectedly requires the UPR transcription factor X-box binding protein 1 (XBP-1). Synergistic IFN-beta induction also occurs in HLA-B27/human beta(2)m-transgenic rat macrophages exhibiting a UPR as a consequence of HLA-B27 up-regulation, where it correlates with activation of XBP-1 splicing. Together these findings indicate that the cellular response to endogenous 'danger' that disrupts ER homeostasis is coupled to IFN-beta induction by XBP-1, which has implications for the immune response and the pathogenesis of diseases involving the UPR.

Smith JA; Turner MJ; DeLay ML; Klenk EI; Sowders DP; Colbert RA

2008-05-01

204

Toxicity assessment of transgenic papaya ringspot virus of 823-2210 line papaya fruits.  

UK PubMed Central (United Kingdom)

The transgenic papaya is a valuable strategy for creating plants resistant to papaya ringspot virus (PRSV) infection and increasing production. This study was further performed to evaluate the comparative toxicity effects of the newly developed transgenic line of the fruits of two backcross transgenic papaya lines (2210 and 823) and one hybrid line (823-2210) and compare to their parent non-transgenic (TN-2) counterparts. The stability analysis of coat protein (CP) of PRSV was investigated using the digestion stability assays in simulated gastric fluid (SGF), simulated intestinal fluid (SIF), and bile salts to detect the CP fragments. Results revealed that the CP fragments were rapidly hydrolyzed in SGF and were undetectable in organs and gastrointestinal contents in rats. For the genotoxicity, three in vitro assays were conducted and exhibited that non-transgenic and backcross transgenic papaya fruits were negative. Moreover, a repeated animal feeding study was conducted by feeding 2 g/kg of body weight (bw) of non-transgenic and backcross transgenic papaya fruits for 28 days in rats. There were no biological or toxicological significances between non-transgenic and backcross transgenic papaya fruits in rats. The results demonstrated that the backcross transgenic papaya fruit can be recognized as an equivalent substitution for traditional papaya in food safety.

Lin HT; Yen GC; Huang TT; Chan LF; Cheng YH; Wu JH; Yeh SD; Wang SY; Liao JW

2013-02-01

205

Transgene method for plant  

UK PubMed Central (United Kingdom)

A transgenic method of plant, it relates a method of plant transgenic, agrobacterium medium transform to embryo just finishing fecundation, this embryo group to seed on individual plant. We filter the seeds to obtain transform element, this technology adapts to corn, soja, paddy and so on, series of fecundation plant which we can manipulate it's germen. Its character is: no organ cultivation, no liable to gene type and experiment condition, high efficiency.

LIU DEPU; YUAN YING; HAO WENYUAN; ZHENG PEIHE; TAN HUA; DONG YINGSHAN

206

Angiotensin-converting enzyme inhibition, but not AT(1) receptor blockade, in the solitary tract nucleus improves baroreflex sensitivity in anesthetized transgenic hypertensive (mRen2)27 rats.  

Science.gov (United States)

Transgenic hypertensive (mRen2)27 rats overexpress the murine Ren2 gene and have impaired baroreflex sensitivity (BRS) for control of the heart rate. Removal of endogenous angiotensin (Ang)-(1-7) tone using a receptor blocker does not further lower BRS. Therefore, we assessed whether blockade of Ang II with a receptor antagonist or combined reduction in Ang II and restoration of endogenous Ang-(1-7) levels with Ang-converting enzyme (ACE) inhibition will improve BRS in these animals. Bilateral solitary tract nucleus (nTS) microinjections of the AT(1) receptor blocker, candesartan (CAN, 24?pmol in 120?nl, n=9), or a peptidic ACE inhibitor, bradykinin (BK) potentiating nonapeptide (Pyr-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro; BPP9?, 9?nmol in 60?nl, n=12), in anesthetized male (mRen2)27 rats (15-25 weeks of age) show that AT(1) receptor blockade had no significant effect on BRS, whereas microinjection of BPP9? improved BRS over 60-120?min. To determine whether Ang-(1-7) or BK contribute to the increase in BRS, separate experiments using the Ang-(1-7) receptor antagonist D-Ala(7)-Ang-(1-7) or the BK antagonist HOE-140 showed that only the Ang-(1-7) receptor blocker completely reversed the BRS improvement. Thus, acute AT(1) blockade is unable to reverse the effects of long-term Ang II overexpression on BRS, whereas ACE inhibition restores BRS over this same time frame. As the BPP9? potentiation of BK actions is a rapid phenomenon, the likely mechanism for the observed delayed increase in BRS is through ACE inhibition and elevation of endogenous Ang-(1-7). PMID:21937997

Isa, Katsunori; Arnold, Amy C; Westwood, Brian M; Chappell, Mark C; Diz, Debra I

2011-09-22

207

Rapid one-step purification of native dimeric ALS-associated human Cu/Zn superoxide dismutase from transgenic rat tissues.  

UK PubMed Central (United Kingdom)

Mutated Cu/Zn superoxide dismutase (SOD1) was the first proven cause of amyotrophic lateral sclerosis (ALS) and was the basis for the first animal model. Many approaches, including transgenic and knock-out animals, cell models, and in vitro studies using recombinant hSOD1 mutants and wild-type, have been employed in an attempt to elucidate the gained toxic function. However, a thorough characterization of the properties of hSOD1 mutants produced in vivo has yet to be carried out, primarily due to the lack of a procedure capable of purifying the enzyme from relevant tissues in a manner that avoids potential artifacts. Here we report a new, one-step purification procedure using a semi-preparative polymeric reversed-phase HPLC system, which yields greater than 99% pure enzyme from the spinal cord, and >95% pure from brain, heart, and kidney. This novel approach for purifying 'in vivo expressed' native dimeric SOD1 will facilitate the determination of the true 'as isolated' properties of the enzyme that is responsible for disease, devoid of any expression system, or harsh purification, artifacts. An important new finding related to the specific activity of human SOD1 (normalized to copper content) is also discussed.

Bhogaraju VK; Levi MS; Reed RL; Crow JP

2010-05-01

208

Contribution of FcRn binding to intestinal uptake of IgG in suckling rat pups and human FcRn-transgenic mice.  

UK PubMed Central (United Kingdom)

Immunoglobulin G (IgG) is transcytosed across intestinal epithelial cells of suckling mammals by the neonatal Fc receptor (FcRn); however, the contribution of FcRn vs. FcRn-independent uptake to serum IgG levels had not been determined in either rat pups or human (h)FcRn-expressing mice (Tg276 and Tg32). In isoflurane-anesthetized rodents, serum levels were determined after regional intestinal delivery of human monoclonal antibodies (hIgG) with either wild-type (WT) Fc sequences or variants engineered for different FcRn binding affinities. Detection of full-length hIgG was by immunoassay; intestinal hFcRn and hIgG localization was by immunocytochemistry. High (?g/ml) serum levels of hIgG were detected after proximal intestinal delivery (0.1-10 mg/kg) in 2-wk-old rats. Human FcRn was visualized in epithelial cells of Tg276 mice, but low serum hIgG levels (<10 ng/ml) were obtained. In rat pups, intraintestinal hIgG1 WT administration resulted in dose-related and saturable uptake, whereas uptake of a low FcRn-binding affinity variant was nonsaturable. There were no differences in hIgG levels from systemic and hepatic portal vein serum samples, and intense hIgG immunostaining was noted in villi enterocytes and within lymphatic lacteal-like vessels. This study demonstrated that FcRn-mediated uptake in rat pups accounted for ~80% of serum hIgG levels and that IgG enters the circulation via the lymph and not the hepatic portal vein. The remaining uptake though the immature intestine is nonreceptor mediated. Intestinal epithelial cell hFcRn expression occurred in Tg276 mice, but receptor-mediated transport of IgG was not observed. The suckling rat pup intestine is a mechanistic model of FcRn-IgG-mediated transcytosis.

Kliwinski C; Cooper PR; Perkinson R; Mabus JR; Tam SH; Wilkinson TM; Giles-Komar J; Scallon B; Powers GD; Hornby PJ

2013-02-01

209

Metabonomics study of transgenic Bacillus thuringiensis rice (T2A-1) meal in a 90-day dietary toxicity study in rats.  

UK PubMed Central (United Kingdom)

Rice is one of the most important staple foods in the world. The Cry2A gene was inserted into the rice genome to help the plant combat insects. As the unintended effects of the genetically modified (GM) organisms are the most important barriers to the promotion of GM organisms, we have carried out a useful exploration to establish a new in vivo evaluation model for genetically modified foods by metabonomics methods. In this study, the rats were fed for 90 days with the GM and NON-GM rice diets. The changes in metabolites of the urine were detected using (1)H-NMR. The metabonomics were analyzed to see whether the GM rice can induce the metabolite changes in the rats' urine when compared with the NON-GM rice group. The multivariate analysis and ANOVA were used to determine the differences and the significance of differences respectively, and eventually we concluded that these differences did not have a biological significance. The conclusion of the metabonomics was comparable with that from the traditional method. As a non-invasive and dynamic monitoring method, metabonomics will be a new way of assessing the food safety of GM foods.

Cao S; Xu W; Luo Y; He X; Yuan Y; Ran W; Liang L; Huang K

2011-07-01

210

Metabonomics study of transgenic Bacillus thuringiensis rice (T2A-1) meal in a 90-day dietary toxicity study in rats.  

Science.gov (United States)

Rice is one of the most important staple foods in the world. The Cry2A gene was inserted into the rice genome to help the plant combat insects. As the unintended effects of the genetically modified (GM) organisms are the most important barriers to the promotion of GM organisms, we have carried out a useful exploration to establish a new in vivo evaluation model for genetically modified foods by metabonomics methods. In this study, the rats were fed for 90 days with the GM and NON-GM rice diets. The changes in metabolites of the urine were detected using (1)H-NMR. The metabonomics were analyzed to see whether the GM rice can induce the metabolite changes in the rats' urine when compared with the NON-GM rice group. The multivariate analysis and ANOVA were used to determine the differences and the significance of differences respectively, and eventually we concluded that these differences did not have a biological significance. The conclusion of the metabonomics was comparable with that from the traditional method. As a non-invasive and dynamic monitoring method, metabonomics will be a new way of assessing the food safety of GM foods. PMID:21594293

Cao, Sishuo; Xu, Wentao; Luo, YunBo; He, Xiaoyun; Yuan, Yanfang; Ran, Wenjun; Liang, Lixing; Huang, Kunlun

2011-05-19

211

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

Science.gov (United States)

Stem cells provide novel sources of cell therapies for motor neuron disease that have recently entered clinical trials. In the present study, we transplanted human neural stem cells (NSCs) into the ventral horn of both the lumbar (L4–L5) and cervical (C4–C5) protuberance of SOD G93A rats, in an effort to test the feasibility and general efficacy of a dual grafting paradigm addressing several muscle groups in the front limbs, hind limbs and the respiratory apparatus. Transplantation was done prior to the onset of motor neuron disease. Compared with animals that had received dead NSC grafts (serving as controls), rats with live NSCs grafted at the two spinal levels lived 17 days longer. Disease onset in dually grafted animals was delayed by 10 days compared to control animals. Disease duration in NSC-grafted animals was longer by 7 days compared to controls. Our results support the potential of NSC grafts at multiple levels of spinal cord as future cellular therapy for motor neuron disease.

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

2011-01-01

212

Generation of transgenic frogs.  

UK PubMed Central (United Kingdom)

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

Loeber J; Pan FC; Pieler T

2009-01-01

213

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

DEFF Research Database (Denmark)

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

van Onna, M; Jurik, A G

2011-01-01

214

Transgenic Crops for Herbicide Resistance  

Science.gov (United States)

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

215

Relative transgene expression frequencies in homozygous versus hemizygous transgenic mice.  

UK PubMed Central (United Kingdom)

We have used a simple binomial model of stochastic transgene inactivation at the level of the chromosome or transgene, rather than the cellular level, for the analysis of two mouse transgenic lines that show variegated patterns of expression. This predicts the percentages of cells that express one, both or neither alleles of the transgene in homozygotes from the observed percentages of cells, which express the transgene in hemizygotes. It adequately explained the relationship between the numbers of cells expressing the transgene in hemizygous and homozygous mosaic 21OH/LacZ mouse adrenals and mosaic BLG/7 mouse mammary glands. The binomial model also predicted that a small proportion of cells in mosaic mammary glands of BLG/7 homozygotes would express both BLG/7 alleles but published data indicated that all cells expressing the transgene showed monoallelic expression. Although it didn't fit all of the BLG/7 data as precisely as a more complex model, which used several ad hoc assumptions to explain these results, the simple binomial model was able to explain the relationship in observed transgene expression frequencies between hemizygous and homozygous mosaic tissues for both 21OH/LacZ and BLG/7 mice. It may prove to be a useful general model for analysing other transgenic animals showing mosaic transgene expression.

Chang SP; Opsahl ML; Whitelaw CB; Morley SD; West JD

2013-07-01

216

Mitochondrial superoxide mediates labile iron level: Evidence from Mn-SOD transgenic mice and heterozygous knockout mice and isolated rat liver mitochondria.  

UK PubMed Central (United Kingdom)

Superoxide is the main reactive oxygen species (ROS) generated by aerobic cells primarily in mitochondria. It is also capable of producing other ROS and reactive nitrogen species (RNS). Moreover, superoxide has the potential to release iron from its protein complexes. Unbound or loosely bound cellular iron, known as labile iron, can catalyze the formation of the highly reactive hydroxyl radical. ROS/RNS can cause mitochondrial dysfunction and damage. Manganese superoxide dismutase (Mn-SOD) is the chief ROS scavenging enzyme and thereby the primary antioxidant involved in protecting mitochondria from oxidative damage. To investigate whether mitochondrial superoxide mediates labile iron in vivo, the levels of labile iron were determined in the tissues of mice overexpressing Mn-SOD and heterozygous Mn-SOD knockout mice. Furthermore, the effect of increased mitochondrial superoxide generation on labile iron levels was determined in isolated rat liver mitochondria exposed to various electron transport inhibitors. The results clearly showed that increased expression of Mn-SOD significantly lowered the levels of labile iron in heart, liver, kidney and skeletal muscle, while decreased expression of Mn-SOD significantly increased the levels of labile iron in the same organs. In addition, the data showed that peroxidative damage to membrane lipids closely correlated with the levels of labile iron in various tissues, and that altering the status of Mn-SOD did not alter the status of other antioxidant systems. Results also showed that increased ROS production in isolated liver mitochondria significantly increased the levels of mitochondrial labile iron. These findings constitute the first evidence which suggest that mitochondrial superoxide is capable of releasing iron from its protein complexes in vivo, and that it could also release iron from protein complexes contained within the organelle.

Ibrahim WH; Habib HM; Kamal H; Clair DK; Chow CK

2013-06-01

217

Calcium electrotransfer for termination of transgene expression in muscle  

DEFF Research Database (Denmark)

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

Hojman, Pernille; Spanggaard, Iben

2011-01-01

218

Transgenic camelina sativa  

UK PubMed Central (United Kingdom)

The present invention relates to plant biotechnology and specifically to a method for genetically transforming Camelina sativa with Agrobacterium-mediated transformation system. It comprises Camelina sativa for producing homologous and heterologous recombinant products including oil and protein products and assessing and screening the efficacy of plant transformation. Also disclosed are transgenic Camelina sativa plants, seeds as well as cells, cell-lines and tissue of Camelina sativa.

KUVSHINOV VIKTOR; KANERVA ANNE; KOIVU KIMMO; KUVSHINOVA SVETLANA; PEHU EIJA

219

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.

Lefesvre Pierre; Attema Joline; van Bekkum Dirk

2003-01-01

220

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.)

1993-01-01

 
 
 
 
221

ALIMENTOS TRANSGÉNICOS TRANSGENIC FOODS  

Directory of Open Access Journals (Sweden)

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

María Soledad Reyes S.; Jaime Rozowski N

2003-01-01

222

ALIMENTOS TRANSGÉNICOS/ TRANSGENIC FOODS  

Scientific Electronic Library Online (English)

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

Reyes S., María Soledad; Rozowski N, Jaime

2003-04-01

223

Overgrowth of skin in growth hormone transgenic mice depends on the presence of male gonads.  

UK PubMed Central (United Kingdom)

Growth hormone has been shown to possess stimulatory effects on various connective tissues. We observed that skin growth in male rat phosphoenolpyruvate carboxykinase-bovine growth hormone transgenic mice (serum growth hormone levels: 740-1940 ng per ml) is progressive with age, resulting in an "oversized coat" phenotype with a marked increase in absolute and relative skin weight and surface area, and in thickness of the dermis. Histologic changes include severe dermal fibrosis and replacement of subdermal adipose tissue by fibrous tissue. Apart from an increase in skin surface area, these changes were not noted in female transgenic mice, arguing for a specific interaction of growth hormone with male sex hormones. To clarify this point, 6 wk old male transgenic mice and control mice were castrated and compared with their noncastrated counterparts in parameters of skin growth at an age of 8 mo. The skin weight of castrated transgenic mice was smaller (p < 0.01) than that of intact transgenic mice both absolutely and relative to body weight. The relative skin weight of castrated transgenic mice was in the same range as in intact and castrated control mice. Absolute and relative skin area of castrated transgenic mice was greater (p < 0. 001 and p < 0.05) than in controls but lower than in intact transgenic mice (p < 0.001 and p < 0.05). When compared with control mice, intact transgenic mice displayed an increase (p < 0.01) in the thickness of dermis. In castrated transgenic mice the thickness of the dermis was in the same range as in control mice. Our findings demonstrate a specific interaction of growth hormone with male sex hormones resulting in a marked stimulation of skin growth.

Wanke R; Milz S; Rieger N; Ogiolda L; Renner-Müller I; Brem G; Hermanns W; Wolf E

1999-12-01

224

Transgenic mice overexpressing aldose reductase in Schwann cells show more severe nerve conduction velocity deficit and oxidative stress under hyperglycemic stress  

Digital Repository Infrastructure Vision for European Research (DRIVER)

To further understand the role of aldose reductase (AR) in the etiology of diabetic neuropathy, we generated transgenic mice that overexpress AR specifically in the Schwann cells under the control of the rat myelin protein zero (P 0) promoter. One of the transgenic mouse lines, which has overexpress...

Song, Z; Fu, DTW; Chan, YS; Leung, S; Chung, SSM; Chung, SK

225

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

226

Transgenic agriculture and environmental indicators  

Directory of Open Access Journals (Sweden)

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

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

2006-01-01

227

[Current status and industrialization of transgenic tomatoes].  

UK PubMed Central (United Kingdom)

In this review, the progress in transgenic tomato research, including disease and insect resistance, herbicide resistance, stress tolerance, long-term storage, quality improvement, and male sterility, were described. The recent researches on producing heterologous proteins using transgenic tomatoes were also reviewed. Furthermore, the industrialization status and problems of transgenic tomatoes were analyzed and the prospects of both research and industrialization in transgenic tomatoes were discussed.

Wang AX; Chen XL

2011-09-01

228

Expression of chicken liver cell adhesion molecule fusion genes in transgenic mice.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The tissue-specific expression of the chicken liver cell adhesion molecule (L-CAM) was studied by generating transgenic mice. The rat insulin II promoter was fused to a chicken L-CAM cDNA or to chicken genomic L-CAM sequences. Mice carrying the cDNA showed no expression of L-CAM. Mice carrying L-CAM...

Begemann, M; Tan, S S; Cunningham, B A; Edelman, G M

229

Transgenic mice produced by retroviral transduction of male germ line stem cells in vivo.  

UK PubMed Central (United Kingdom)

Spermatogonial stem cells are the only stem cells in the postnatal body that can transmit parental genetic information to the offspring, making them an attractive target cell population for animal transgenesis. Although transgenic mice and rats were recently produced by retrovirus transduction of these cells in vitro, with transplantation of the transduced cells into infertile recipients, the difficulty of restoring fertility and preparing recipients using spermatogonial transplantation limits practical application of the technique. In this article, we describe a novel approach for producing transgenic animals by transducing spermatogonial stem cells in vivo using a retrovirus vector. Microinjection of retrovirus into immature seminiferous tubules resulted in the direct transduction of spermatogonial stem cells in situ, and the animals produced transgenic offspring after mating with females. Transgenic mice were produced in C57BL/6, BALB/C, A, and C3H backgrounds, with an average efficiency of 2.8%. The transgene was transmitted stably and expressed in the next generation. The technique overcomes the drawback of the in vitro-transduction approach, and will be useful as a novel method for producing transgenic animals as well as providing a means for analyzing the self-renewal and differentiation processes of spermatogonial stem cells in vivo.

Kanatsu-Shinohara M; Toyokuni S; Shinohara T

2004-10-01

230

Strategies for designing transgenic DNA constructs.  

Science.gov (United States)

Generation and characterization of transgenic mice are important elements of biomedical research. In recent years, transgenic technology has become more versatile and sophisticated, mainly because of the incorporation of recombinase-mediated conditional expression and targeted insertion, site-specific endonuclease-mediated genome editing, siRNA-mediated gene knockdown, various inducible gene expression systems, and fluorescent protein marking and tracking techniques. Site-specific recombinases (such as PhiC31) and engineered endonucleases (such as ZFN and Talen) have significantly enhanced our ability to target transgenes into specific genomic loci, but currently a great majority of transgenic mouse lines are continuingly being created using the conventional random insertion method. A major challenge for using this conventional method is that the genomic environment at the integration site has a substantial influence on the expression of the transgene. Although our understanding of such chromosomal position effects and our means to combat them are still primitive, adhering to some general guidelines can significantly increase the odds of successful transgene expression. This chapter first discusses the major problems associated with transgene expression, and then describes some of the principles for using plasmid and bacterial artificial chromosomes (BACs) for generating transgenic constructs. Finally, the strategies for conducting each of the major types of transgenic research are discussed, including gene overexpression, promoter characterization, cell-lineage tracing, mutant complementation, expression of double or multiple transgenes, siRNA knockdown, and conditional and inducible systems. PMID:23912987

Liu, Chengyu

2013-01-01

231

Strategies for designing transgenic DNA constructs.  

UK PubMed Central (United Kingdom)

Generation and characterization of transgenic mice are important elements of biomedical research. In recent years, transgenic technology has become more versatile and sophisticated, mainly because of the incorporation of recombinase-mediated conditional expression and targeted insertion, site-specific endonuclease-mediated genome editing, siRNA-mediated gene knockdown, various inducible gene expression systems, and fluorescent protein marking and tracking techniques. Site-specific recombinases (such as PhiC31) and engineered endonucleases (such as ZFN and Talen) have significantly enhanced our ability to target transgenes into specific genomic loci, but currently a great majority of transgenic mouse lines are continuingly being created using the conventional random insertion method. A major challenge for using this conventional method is that the genomic environment at the integration site has a substantial influence on the expression of the transgene. Although our understanding of such chromosomal position effects and our means to combat them are still primitive, adhering to some general guidelines can significantly increase the odds of successful transgene expression. This chapter first discusses the major problems associated with transgene expression, and then describes some of the principles for using plasmid and bacterial artificial chromosomes (BACs) for generating transgenic constructs. Finally, the strategies for conducting each of the major types of transgenic research are discussed, including gene overexpression, promoter characterization, cell-lineage tracing, mutant complementation, expression of double or multiple transgenes, siRNA knockdown, and conditional and inducible systems.

Liu C

2013-01-01

232

Transgenic trees and forestry biosafety  

Scientific Electronic Library Online (English)

Full Text Available 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 genetically 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 (more) 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

2006-06-01

233

Transgenic mouse models for ADHD.  

Science.gov (United States)

Attention-deficit hyperactivity disorder (ADHD) is a developmental disorder characterized by symptoms of inattention, impulsivity and hyperactivity that adversely affect many aspects of life. Whereas the etiology of ADHD remains unknown, growing evidence indicates a genetic involvement in the development of this disorder. The brain circuits associated with ADHD are rich in monoamines, which are involved in the mechanism of action of psychostimulants and other medications used to treat this disorder. Dopamine (DA) is believed to play a major role in ADHD but other neurotransmitters are certainly also involved. Genetically modified mice have become an indispensable tool used to analyze the contribution of genetic factors in the pathogenesis of human disorders. Although rodent models cannot fully recapitulate complex human psychiatric disorders such as ADHD, transgenic mice offer an opportunity to directly investigate in vivo the specific roles of novel candidate genes identified in ADHD patients. Several knock-out and transgenic mouse models have been proposed as ADHD models, mostly based on targeting genes involved in DA transmission, including the gene encoding the dopamine transporter (DAT1). These mutant models provided an opportunity to evaluate the contribution of dopamine-related processes to brain pathology, to dissect the neuronal circuitry and molecular mechanisms involved in the antihyperkinetic action of psychostimulants and to evaluate novel treatments for ADHD. New transgenic models mouse models targeting other genes have recently been proposed for ADHD. Here, we discuss the recent advances and pitfalls in modeling ADHD endophenotypes in genetically altered animals. PMID:23681253

Leo, Damiana; Gainetdinov, Raul R

2013-05-17

234

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

2012-11-29

235

Insect-resistant transgenic Pinus radiata.  

UK PubMed Central (United Kingdom)

Transgenic radiata pine (Pinus radiata D. Don) plants containing a Bacillus thuringiensis (Bt) toxin gene, crylAc, were produced by means of biolistic transformation of embryogenic tissue. Using the selectable marker gene nptII and corresponding geneticin selection, 20 independent transgenic lines from five genotypes were established. Over 200 plants regenerated from ten transgenic lines were successfully transferred to soil. The integration and expression of the introduced genes in transgenic tissue and/or plants were confirmed by PCR, Southern hybridisation and neomycin phosphotransferase II (NPTII) and Bt ELISA assays. Bioassays with larvae of the painted apple moth, Teia anartoides, demonstrated that transgenic plants displayed variable levels of resistance to insect damage, with one transgenic line being highly resistant to feeding damage.

Grace LJ; Charity JA; Gresham B; Kay N; Walter C

2005-05-01

236

Insect-resistant transgenic Pinus radiata.  

Science.gov (United States)

Transgenic radiata pine (Pinus radiata D. Don) plants containing a Bacillus thuringiensis (Bt) toxin gene, crylAc, were produced by means of biolistic transformation of embryogenic tissue. Using the selectable marker gene nptII and corresponding geneticin selection, 20 independent transgenic lines from five genotypes were established. Over 200 plants regenerated from ten transgenic lines were successfully transferred to soil. The integration and expression of the introduced genes in transgenic tissue and/or plants were confirmed by PCR, Southern hybridisation and neomycin phosphotransferase II (NPTII) and Bt ELISA assays. Bioassays with larvae of the painted apple moth, Teia anartoides, demonstrated that transgenic plants displayed variable levels of resistance to insect damage, with one transgenic line being highly resistant to feeding damage. PMID:15668791

Grace, Lynette J; Charity, Julia A; Gresham, Belinda; Kay, Nod; Walter, Christian

2005-01-25

237

Transgene integration, organization and interaction in plants.  

UK PubMed Central (United Kingdom)

It has been appreciated for many years that the structure of a transgene locus can have a major influence on the level and stability of transgene expression. Until recently, however, it has been common practice to discard plant lines with poor or unstable expression levels in favor of those with practical uses. In the last few years, an increasing number of experiments have been carried out with the primary aim of characterizing transgene loci and studying the fundamental links between locus structure and expression. Cereals have been at the forefront of this research because molecular, genetic and cytogenetic analysis can be carried out in parallel to examine transgene loci in detail. This review discusses what is known about the structure and organization of transgene loci in cereals, both at the molecular and cytogenetic levels. In the latter case, important links are beginning to be revealed between higher order locus organization, nuclear architecture, chromatin structure and transgene expression.

Kohli A; Twyman RM; Abranches R; Wegel E; Stoger E; Christou P

2003-05-01

238

Determining gene flow in transgenic cotton.  

UK PubMed Central (United Kingdom)

Gene flow is one of the major concerns associated with the release of transgenic plants into the environment. Unrestricted gene flow can results in super weeds, reduction in species fitness and genetic diversity, and contamination of traditional plants and foods. Thus, it is important and also necessary to evaluate the extent of gene flow in the field for transgenic plants already released or being considered for a release. Transgenic cotton is among the first transgenic crops for commercialization, which are widely cultivated around the world. In this chapter, we use transgenic insect resistant cotton and herbicide-tolerant cotton as two examples to present a field practice method for determining transgene flow in cotton. The procedure includes three major sections: (1) field design, (2) seed collection, and (3) field and lab bioassay.

Pan X

2013-01-01

239

Derivation of a germline competent transgenic Fischer 344 embryonic stem cell line.  

UK PubMed Central (United Kingdom)

Embryonic stem (ES) cell-based gene manipulation is an effective method for the generation of mutant animal models in mice and rats. Availability of germline-competent ES cell lines from inbred rat strains would allow for creation of new genetically modified models in the desired genetic background. Fischer344 (F344) males carrying an enhanced green fluorescence protein (EGFP) transgene were used as the founder animals for the derivation of ES cell lines. After establishment of ES cell lines, rigorous quality control testing that included assessment of pluripotency factor expression, karyotype analysis, and pathogen/sterility testing was conducted in selected ES cell lines. One male ES cell line, F344-Tg.EC4011, was further evaluated for germline competence by injection into Dark Agouti (DA) X Sprague Dawley (SD) blastocysts. Resulting chimeric animals were bred with wild-type SD mates and germline transmissibility of the ES cell line was confirmed by identification of pups carrying the ES cell line-derived EGFP transgene. This is the first report of a germline competent F344 ES cell line. The availability of a new germline competent ES cell line with a stable fluorescence reporter from an inbred transgenic rat strain provides an important new resource for genetic manipulations to create new rat models.

Men H; Bryda EC

2013-01-01

240

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

Science.gov (United States)

Embryonic stem (ES) cell-based gene manipulation is an effective method for the generation of mutant animal models in mice and rats. Availability of germline-competent ES cell lines from inbred rat strains would allow for creation of new genetically modified models in the desired genetic background. Fischer344 (F344) males carrying an enhanced green fluorescence protein (EGFP) transgene were used as the founder animals for the derivation of ES cell lines. After establishment of ES cell lines, rigorous quality control testing that included assessment of pluripotency factor expression, karyotype analysis, and pathogen/sterility testing was conducted in selected ES cell lines. One male ES cell line, F344-Tg.EC4011, was further evaluated for germline competence by injection into Dark Agouti (DA) X Sprague Dawley (SD) blastocysts. Resulting chimeric animals were bred with wild-type SD mates and germline transmissibility of the ES cell line was confirmed by identification of pups carrying the ES cell line-derived EGFP transgene. This is the first report of a germline competent F344 ES cell line. The availability of a new germline competent ES cell line with a stable fluorescence reporter from an inbred transgenic rat strain provides an important new resource for genetic manipulations to create new rat models.

Men, Hongsheng; Bryda, Elizabeth C.

2013-01-01

 
 
 
 
241

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

242

TRANSGENIC PLANTS EXPRESSING A VIRAL ANTIFUNGAL PROTEIN  

UK PubMed Central (United Kingdom)

Transgenic plants expressing the KP4 antifungal protein are provided which exhibit high levels of antifungal resistance. Such transgenic plants contain a recombinant DNA construct comprising a heterologous signal peptide sequence that is operably linked to a non-native nucleic acid sequence encoding a mature KP4 antifungal protein.

SMITH THOMAS; SHAH DILIP MAGANLAL

243

THE USE OF TRANSGENES FOR WEED MANAGEMENT  

Science.gov (United States)

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

244

[New advances in animal transgenic technology].  

UK PubMed Central (United Kingdom)

Animal transgenic technology is one of the fastest growing biotechnology in the 21st century. It is used to integrate foreign genes into the animal genome by genetic engineering technology so that foreign genes can be expressed and inherited to the offspring. The transgenic efficiency and precise control of gene expression are the key limiting factors on preparation of transgenic animals. A variety of transgenic techniques are available, each of which has its own advantages and disadvantages and still needs further study because of unresolved technical and safety issues. With the in-depth research, the transgenic technology will have broad application prospects in the fields of exploration of gene function, animal genetic improvement, bioreactor, animal disease models, organ transplantation and so on. This article reviews the recently developed animal gene transfer techniques, including germline stem cell mediated method to improve the efficiency, gene targeting to improve the accuracy, RNA interference (RNAi)-mediated gene silencing technology, and the induced pluripotent stem cells (iPS) transgenic technology. The new transgenic techniques can provide a better platform for the study of trans-genic animals and promote the development of medical sciences, livestock production, and other fields.

Sun ZH; Miao XY; Zhu RL

2010-06-01

245

The Demonstration of Transgenic Tobacco  

UK PubMed Central (United Kingdom)

transformed regeneration plants derived from three varieties of tobacoo whic h were transformed by the leaf-method and Kanamycin selection were tested throug h PCR electrophoresis assay, under the media of plasmoid PBLGC (PBLGC contained chitinase gene and #beta#-1, 3-glucanase gene and NTP-II gene) by using promtor 35S, chitinase gene and #beta#-1, 3-glucanase gene primer separately. 92,72 and 47 posi tive plants were obtained respectively. There were 36 positive plants containing both chitinase gene and #beta#-1, 3-glucanase gene. Parts of transgenic plants were tested by PCR-Southern. The result showed that the transferring frequency of foreign gene was related to plant variety and the length of foreign gene. The prog enies of four transgenic tobacco were demonstrated by PCR and PCR-Southern hybri dization analysis of tobacco DNAs. The result indicates that chitinase gene and #beta#-1, 3-glucanase gene had stably inherited to progenies, but their rates of inh eritance were different.

Zhang Yanni; Qu Min; Zhuo Lihuan; Xu Xiangling; Yang Liping

2003-01-01

246

THERMAL SOFTENING OF TRANSGENIC ASPEN  

Directory of Open Access Journals (Sweden)

Full Text Available Studies on the softening behavior of in situ lignin of normal wood in a given species have never been performed before due to the relatively narrow lignin content and lignin structural variation within one species. Using transgenic trees with different levels of lignin content and/or syringyl to guaiacyl propane (S/G) ratio helped us to overcome this problem. Submersion three-point bending and parallel-plate compression-torsion dynamic mechanical analyses were conducted on one-year-old wild type and transgenic aspen (Populus tremuloides Michx.). The different genetic modifications included groups with reduced lignin content, increased S/G ratio, and both reduced lignin content and increased S/G ratio. Measurements with both methods revealed a statistically significant decrease in glass transition temperature in the reduced-lignin genetic group compared to the wild-type. Increase in the S/G ratio did not affect the thermo-mechanical properties; these results contradict claims that increasing the methoxyl groups would reduce lignin cross-linking and the glass transition temperature.

Bbalazs Horvath; Perry Peralta; Charles Frazier; Ilona Maria Peszlen Mail

2011-01-01

247

Wheat Storage Proteins in Transgenic Rice Endosperm.  

UK PubMed Central (United Kingdom)

A wheat HMW glutenin subunit (HMW-GS) protein in transgenic rice expressing was characterised to study its effects on the protein size distribution and on the protein expression pattern of the rice endosperm; as well as on 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 non-transgenic plants, combined with proteomic based studies, indicated increased 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 positive effect on the functional properties of rice dough by significantly increasing the size distribution of the polymeric protein fraction and modify the dough mixing parameters.

Oszvald M; Balázs G; Pólya S; Tömösközi S; Appels R; Bekes F; Tamas L

2013-06-01

248

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.

2007-01-01

249

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

Energy Technology Data Exchange (ETDEWEB)

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.

Tae, Seong Ho; Min, Jung Joon [Chonnam National University Medical School, Gwangju (Korea, Republic of); Le, Uyenchi N.; Padmanabhan, Parasuraman [Singapore Bio-Imaging Imaging Consortium, Singapore (Singapore)

2007-07-01

250

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

UK PubMed Central (United Kingdom)

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

Luna AS; da Silva AP; Pinho JS; Ferré J; Boqué R

2013-01-01

251

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

Science.gov (United States)

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

252

Expression of multiple proteins in transgenic plants  

Energy Technology Data Exchange (ETDEWEB)

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

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

2002-01-01

253

[Construction of Runx1 transgenic mice].  

UK PubMed Central (United Kingdom)

This study was aimed to construct transgenic mouse model with target for Runxl gene. Runxl cDNA of mice was amplified by PCR from pcDNA3. 1 Flag Runx1 FL vector and inserted into ptetO7-Asc-IRES-EGFP vector to form a recombinant vector, and then the recombinant vector was injected into fertilized egg by microinjection technology to get a transgenic mouse. The results of PCR and Southern blot indicated that the Runx1 transgenic mouse was constructed successfully, and this could provide an important tool for studying the function of Runxl gene in vivo.

Tang X; Li J; Liu X; Lei Y; Su Q; Wu Q; Luo F

2013-06-01

254

Transgene insertion pattern analysis using genomic walking in a transgenic mouse line.  

UK PubMed Central (United Kingdom)

A transgene mapping technique (Noguchi et al., Exp. Anim. 53:103-111, 2004) is described that can be used to analyze transgene integration patterns in transgenic mice. The technique was used to reveal that a transgenic mouse line (GM1-sy#116) harbored inverted and direct tandem repeats of both intact and partial pCAGGS-based transgenes in the G2 region of chromosome 1. This complicated concatenation of transgenes may have been caused by simple end-joining of DNA constructs fragmented by exposure to UV transillumination during gel-purification, and by nuclease digestion inside zygote pronuclei. The results suggest that care should be taken to avoid unwanted fragmentation during the preparation of vector constructs.

Suzuki O; Hata T; Takekawa N; Koura M; Takano K; Yamamoto Y; Noguchi Y; Uchio-Yamada K; Matsuda J

2006-01-01

255

Transgene insertion pattern analysis using genomic walking in a transgenic mouse line.  

Science.gov (United States)

A transgene mapping technique (Noguchi et al., Exp. Anim. 53:103-111, 2004) is described that can be used to analyze transgene integration patterns in transgenic mice. The technique was used to reveal that a transgenic mouse line (GM1-sy#116) harbored inverted and direct tandem repeats of both intact and partial pCAGGS-based transgenes in the G2 region of chromosome 1. This complicated concatenation of transgenes may have been caused by simple end-joining of DNA constructs fragmented by exposure to UV transillumination during gel-purification, and by nuclease digestion inside zygote pronuclei. The results suggest that care should be taken to avoid unwanted fragmentation during the preparation of vector constructs. PMID:16508214

Suzuki, Osamu; Hata, Tomoko; Takekawa, Naho; Koura, Minako; Takano, Kaoru; Yamamoto, Yoshie; Noguchi, Yoko; Uchio-Yamada, Kozue; Matsuda, Junichiro

2006-01-01

256

Clinical chemistry of human FcRn transgenic mice.  

UK PubMed Central (United Kingdom)

Mice genetically engineered to express human FcRn are valuable models for the evaluation of therapeutic antibodies in the context of human FcRn in vivo. However, only limited clinical chemistry information on these mouse strains is available. Thus, we have compared 30 clinical chemical parameters of C57BL/6J wild-type mice, murine FcRn-knockout mice, and two human FcRn transgenic mouse strains expressing human FcRn in the absence of murine FcRn. Since FcRn-mediated recycling prevents albumin and IgG from intracellular degradation, significant differences for both proteins were observed in the murine FcRn-knockout mice. Mice lacking FcRn show lower IgG and albumin levels compared to wild-type mice. The most prominent differences in clinical chemical parameters can be explained by secondary effects of the altered albumin levels of murine FcRn-knockout mice on liver metabolism, as similar tendencies have been observed in analbuminemic Nagase rats and hypoalbuminemic human patients, showing an overall increased liver metabolism. Both human FcRn transgenic strains show clinical chemical parameters similar to those found for wild-type mice, with the exception of endogenous IgG levels, which are greatly reduced in these mice.

Stein C; Kling L; Proetzel G; Roopenian DC; de Angelis MH; Wolf E; Rathkolb B

2012-04-01

257

Ethics and Transgenic Crops: a Review  

Scientific Electronic Library Online (English)

Full Text Available Abstract in english This article represents a review of some of the ethical dilemmas that have arisen as a result of the development and deployment of transgenic crop plants. The potential for transgenic crops to alleviate human hunger and the possible effects on human health are discussed. Risks and benefits to the environment resulting from genetic engineering of crops for resistance to biotic and abiotic stresses are considered, in addition to effects on biodiversity. The socio-economic i (more) mpacts and distribution of benefits from transgenic technologies are reviewed. Fundamental issues of man?s relationship with nature and the environment, and theological matters are also addressed. An almost unprecedented amount of discussion has been stimulated on the merits and demerits of genetic engineering of crop plants, and has divided both the public and scientific communities. The arguments for and against transgenics are invariably based on visions of the new technology from widely different ethical perspectives.

Robinson, Jonathan

1999-08-01

258

Transgenic Saccharomyces Cerevisiae and Method for Bioremediation.  

Science.gov (United States)

An isolated and purified transgenic Saccharomyces cerevisiae yeast cell comprising a disrupted ACR3 gene and an isolated DNA sequence comprising a promoter operably linked to a nucleic acid molecule encoding yeast cadmium factor resistance protein Ycf1p, ...

B. Rosen M. Ghosh

2004-01-01

259

Evaluating genetic containment strategies for transgenic plants.  

Science.gov (United States)

One of the primary concerns about genetically engineered crop plants is that they will hybridize with wild relatives, permitting the transgene to escape into the environment. The likelihood that a transgene will spread in the environment depends on its potential fitness impact. The fitness conferred by various transgenes to crop and/or wild-type hybrids has been evaluated in several species. Different strategies have been developed for reducing the probability and impact of gene flow, including physical separation from wild relatives and genetic engineering. Mathematical models and empirical experimental evidence suggest that genetic approaches have the potential to effectively prevent transgenes from incorporating into wild relatives and becoming established in wild populations that are not reproductively isolated from genetically engineered crops. PMID:16460821

Lee, David; Natesan, Ellen

2006-02-07

260

Evaluating genetic containment strategies for transgenic plants.  

UK PubMed Central (United Kingdom)

One of the primary concerns about genetically engineered crop plants is that they will hybridize with wild relatives, permitting the transgene to escape into the environment. The likelihood that a transgene will spread in the environment depends on its potential fitness impact. The fitness conferred by various transgenes to crop and/or wild-type hybrids has been evaluated in several species. Different strategies have been developed for reducing the probability and impact of gene flow, including physical separation from wild relatives and genetic engineering. Mathematical models and empirical experimental evidence suggest that genetic approaches have the potential to effectively prevent transgenes from incorporating into wild relatives and becoming established in wild populations that are not reproductively isolated from genetically engineered crops.

Lee D; Natesan E

2006-03-01

 
 
 
 
261

AN APPROACH TO TRANSGENIC CROP MONITORING  

Science.gov (United States)

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

262

Growth differentiation factor-11 transgenic mice  

UK PubMed Central (United Kingdom)

A transgenic mouse whose genome comprises a disruption of the endogenous growth differentiation factor-11 (GDF-11) gene is disclosed. Also disclosed are methods for making such mice. The mice exhibit a phenotype of increased muscle tissue.

LEE SE-JIN; MCPHERRON ALEXANDRA C

263

[Detection of transgenic crop with gene chip].  

Science.gov (United States)

Some selected available sequences of reporter genes,resistant genes, promoters and terminators are amplified by PCR for the probes of transgenic crop detection gene chip. These probes are arrayed at definite density and printed on the surface of amino-slides by bioRobot MicroGrid II. Results showed that gene chip worked quickly and correctly, when transgenic rice, pawpaw,maize and soybean were applied. PMID:15639876

Huang, Ying-Chun; Sun, Chun-Yun; Feng, Hong; Hu, Xiao-Dong; Yin, Hai-Bin

2003-05-01

264

Improvement of lysine content in transgenic tobacco  

UK PubMed Central (United Kingdom)

In this experiment, the lysine-rich protein gene was transformed into tobacco by leaf disc via Agrobacterium tumefaciens LBA4404 as a vehicle. Histochemical staining for GUS activity, PCR and Southern blot demonstrated that lys was integrated into the genome of tobacco plants. Data analysis showed that lysine content in most of 10 transgenic plants was obviously improved and the content in 1 transgenic plant was increased by 49.89%.

Wang Yiqun; Zheng Jingui; Xie Baogui

2005-01-01

265

Transgenic animals and their application in medicine  

Directory of Open Access Journals (Sweden)

Full Text Available Transgenic animals are animals that are genetically altered to have traits that mimic symptoms of specific human pathologies. They provide genetic models of various human diseases which are important in understanding disease and developing new targets. In early 1980 Gordon and co-workers described the first gene addition experiment using the microinjection technology and since then the impact of transgenic technology on basic research has been significant. Within 20 years of its inception, ATryn the first drug approved by USFDA from transgenic animals was developed and it has opened door to drugs from transgenic animals. In addition, they are looked upon as potential future donors for xenotransplantation. With increasing knowledge about the genetics and improvements in the transgenetic technology numerous useful applications like biologically safe new-generation drugs based on human regulatory proteins are being developed.Various aspects of concern in the coming years are the regulatory guidelines, ethical issues and patents related to the use of transgenic animals. This modern medicine is on the threshold of a pharmacological revolution. Use of transgenic animals will provide solutions for drug research, xenotransplantation, clinical trials and will prove to be a new insight in drug development.

Bagle TR, Kunkulol RR, Baig MS, More SY

2013-01-01

266

TRANSGENIC FISH MODEL IN ENVIRONMENTAL TOXICOLOGY  

Directory of Open Access Journals (Sweden)

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

Madhuri Sharma; A.K. Mandloi; Govind Pandey; A.B. Shrivastav

2012-01-01

267

Selenoprotein-transgenic Chlamydomonas reinhardtii.  

UK PubMed Central (United Kingdom)

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.

Hou Q; Qiu S; Liu Q; Tian J; Hu Z; Ni J

2013-03-01

268

Selenoprotein-Transgenic Chlamydomonas reinhardtii  

Directory of Open Access Journals (Sweden)

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.

Qintang Hou; Shi Qiu; Qiong Liu; Jing Tian; Zhangli Hu; Jiazuan Ni

2013-01-01

269

Transgenic pigs carrying both hHO-1 and hDAF transgenes for xenotransplantation  

UK PubMed Central (United Kingdom)

The invention provides a transgenic animal carrying two transgenes, one encoding a human decay accelerating factor (hDAF) and the other encoding a human heme oxygenase-1 (hHO)-1, which are useful for providing cells, tissues or organs therefrom for xenotransplantation.

TU CHING-FU; YANG CHI-KAI; LIU MING-SHING; HO LIN-LIN; HUANG KUEI-FEN; LEE CHUN-JEAN; TAI HAO-CHIH; HUANG KUEI-FENG

270

Transgenic cotton: from biotransformation methods to agricultural application.  

UK PubMed Central (United Kingdom)

Transgenic cotton is among the first transgenic plants commercially adopted around the world. Since it was first introduced into the field in the middle of 1990s, transgenic cotton has been quickly adopted by cotton farmers in many developed and developing countries. Transgenic cotton has offered many important environmental, social, and economic benefits, including reduced usage of pesticides, indirect increase of yield, minimizing environmental pollution, and reducing labor and cost. Agrobacterium-mediated genetic transformation method is the major method for obtaining transgenic cotton. However, pollen tube pathway-mediated method is also used, particularly by scientists in China, to breed commercial transgenic cotton. Although transgenic cotton plants with disease-resistance, abiotic stress tolerance, and improved fiber quality have been developed in the past decades, insect-resistant and herbicide-tolerant cotton are the two dominant transgenic cottons in the transgenic cotton market.

Zhang B

2013-01-01

271

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

UK PubMed Central (United Kingdom)

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

Jen MC; Baler K; Hood AR; Shin S; Shea LD; Ameer GA

2013-05-01

272

Lessons in obesity from transgenic animals.  

UK PubMed Central (United Kingdom)

Many genetic manipulations have created models of obesity, leanness or resistance to dietary obesity in mice, often providing insights into molecular mechanisms that affect energy balance, and new targets for anti-obesity drugs. Since many genes can affect energy balance in mice, polymorphisms in many genes may also contribute to obesity in humans, and there may be many causes of primary leptin resistance. Secondary leptin resistance (due to high leptin levels) can be investigated by combining the ob mutation with other obesity genes. Some transgenic mice have failed to display the expected phenotype, or have even been obese when leanness was expected. Compensatory changes in the expression of other genes during development, or opposing influences of the gene on energy balance, especially in global knockout mice, may offer explanations for such findings. Obesity has been separated from insulin resistance in some transgenic strains, providing new insights into the mechanisms that usually link these phenotypes. It has also been shown that in some transgenic mice, obesity develops without hyperphagia, or leanness without hypophagia, demonstrating that generalised physiological explanations for obesity in individual humans may be inappropriate. Possibly the most important transgenic model of obesity so far created is the Type 1 11beta-hydroxysteroid dehydrogenase over-expressing mouse, since this models the metabolic syndrome in humans. The perspectives into obesity offered by transgenic mouse models should assist clinical researchers in the design and interpretation of their studies in human obesity.

Arch JR

2002-11-01

273

Lessons in obesity from transgenic animals.  

Science.gov (United States)

Many genetic manipulations have created models of obesity, leanness or resistance to dietary obesity in mice, often providing insights into molecular mechanisms that affect energy balance, and new targets for anti-obesity drugs. Since many genes can affect energy balance in mice, polymorphisms in many genes may also contribute to obesity in humans, and there may be many causes of primary leptin resistance. Secondary leptin resistance (due to high leptin levels) can be investigated by combining the ob mutation with other obesity genes. Some transgenic mice have failed to display the expected phenotype, or have even been obese when leanness was expected. Compensatory changes in the expression of other genes during development, or opposing influences of the gene on energy balance, especially in global knockout mice, may offer explanations for such findings. Obesity has been separated from insulin resistance in some transgenic strains, providing new insights into the mechanisms that usually link these phenotypes. It has also been shown that in some transgenic mice, obesity develops without hyperphagia, or leanness without hypophagia, demonstrating that generalised physiological explanations for obesity in individual humans may be inappropriate. Possibly the most important transgenic model of obesity so far created is the Type 1 11beta-hydroxysteroid dehydrogenase over-expressing mouse, since this models the metabolic syndrome in humans. The perspectives into obesity offered by transgenic mouse models should assist clinical researchers in the design and interpretation of their studies in human obesity. PMID:12508949

Arch, J R S

2002-11-01

274

BAC Manipulations for Making BAC Transgene Arrays.  

UK PubMed Central (United Kingdom)

Chromosome tagging using lac or tet operator repeats for in vivo visualization of chromosome dynamics has now become a standard methodology used in a range of organisms. One variation of this approach has been to build transgene arrays creating artificial chromosome blocks to study various aspects of chromatin structure, transcription, replication, or DNA repair. Previously, plasmid transgenes with or without subsequent gene amplification have been used to build these arrays. However, plasmid arrays typically show heterochromatic properties, while gene amplification typically results in chromosome instability of the amplified regions. To avoid these problems, we are now building transgene arrays from large genomic DNA inserts cloned in bacterial artificial chromosomes (BAC). These BAC transgenes show transcriptional levels within several fold of endogenous genes while also exhibiting targeting to specific nuclear compartments similar to the targeting of the endogenous genes. Here we describe Tn5 transposition and BAC recombineering methods used to retrofit BACs for their use in building BAC transgene arrays. This includes insertion of operator repeats and selectable markers into these BACs as well as targeted insertion or deletion of BAC sequences.

Khanna N; Bian Q; Plutz M; Belmont AS

2013-01-01

275

Growth factor transgenes interactively regulate articular chondrocytes.  

UK PubMed Central (United Kingdom)

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.

Shi S; Mercer S; Eckert GJ; Trippel SB

2013-04-01

276

Gene stability in transgenic aspen (Populus). II. Molecular characterization of variable expression of transgene in wild and hybrid aspen.  

Science.gov (United States)

In many annual plant species, transgene inactivation occurs most often when multiple incomplete/complete copies of the transgene are present in a genome. The expression of single-copy transgene loci may also be negatively influenced by the flanking plant DNA and/or chromosomal location (position effect). To understand transgene silencing in a long-lived tree system, we analyzed several wild (Populus tremula L.) and hybrid (P. tremula L. x P. tremuloides Michx.) aspen lines transgenic to the rolC phenotypical marker system and grown under in vitro, greenhouse and field conditions. The morphological features of the 35S-rolC gene construct were used to screen lines with altered transgene expression, which was later confirmed by Northern experiments. Molecular analyses of hybrid aspen revealed that transgene inactivation was always a consequence of transgene repeats. In wild non-hybrid aspen, however, multiple-insertion-based altered or loss of rolC expression was observed only in three out of six lines showing transgene inactivation. Sequencing analysis revealed AT-rich patches at the transgene flanking genomic regions of some of the wild aspen transgenic lines. One wild aspen line showing variable rolC expression revealed characteristic integration of the transgene into genomic regions containing a high AT content (85% or more). In the remaining two wild aspen transgenic lines unstable for rolC expression, single-copy integration and non-AT-rich or repeat-free transgene flanking regions were found. A partial suppression of rolC was observed in some plants of one of the field-grown wild aspen transgenic lines. In the other wild aspen transgenic line an additional mutant phenotype along with transgene inactivation was found. This indicates that the host genome has some control over expression of a transgene, and the possible role of AT-rich regions in defense against foreign DNA. PMID:11678277

Kumar, S; Fladung, M

2001-09-01

277

Gene stability in transgenic aspen (Populus). II. Molecular characterization of variable expression of transgene in wild and hybrid aspen.  

UK PubMed Central (United Kingdom)

In many annual plant species, transgene inactivation occurs most often when multiple incomplete/complete copies of the transgene are present in a genome. The expression of single-copy transgene loci may also be negatively influenced by the flanking plant DNA and/or chromosomal location (position effect). To understand transgene silencing in a long-lived tree system, we analyzed several wild (Populus tremula L.) and hybrid (P. tremula L. x P. tremuloides Michx.) aspen lines transgenic to the rolC phenotypical marker system and grown under in vitro, greenhouse and field conditions. The morphological features of the 35S-rolC gene construct were used to screen lines with altered transgene expression, which was later confirmed by Northern experiments. Molecular analyses of hybrid aspen revealed that transgene inactivation was always a consequence of transgene repeats. In wild non-hybrid aspen, however, multiple-insertion-based altered or loss of rolC expression was observed only in three out of six lines showing transgene inactivation. Sequencing analysis revealed AT-rich patches at the transgene flanking genomic regions of some of the wild aspen transgenic lines. One wild aspen line showing variable rolC expression revealed characteristic integration of the transgene into genomic regions containing a high AT content (85% or more). In the remaining two wild aspen transgenic lines unstable for rolC expression, single-copy integration and non-AT-rich or repeat-free transgene flanking regions were found. A partial suppression of rolC was observed in some plants of one of the field-grown wild aspen transgenic lines. In the other wild aspen transgenic line an additional mutant phenotype along with transgene inactivation was found. This indicates that the host genome has some control over expression of a transgene, and the possible role of AT-rich regions in defense against foreign DNA.

Kumar S; Fladung M

2001-09-01

278

Toxins for Transgenic Resistance to Hemipteran Pests  

Directory of Open Access Journals (Sweden)

Full Text Available 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 plant protection against some insect pests, Bt toxins exhibit little toxicity against sap sucking insects. Indeed, the pest status of some Hemiptera on Bt-transgenic plants has increased in the absence of pesticide application. The increased pest status of numerous hemipteran species, combined with increased prevalence of resistance to chemical insecticides, provides impetus for the development of biologically based, alternative management strategies. Here, we provide an overview of approaches toward transgenic resistance to hemipteran pests.

Nanasaheb P. Chougule; Bryony C. Bonning

2012-01-01

279

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

280

Generation of transgenic non-human primates with germline transmission.  

UK PubMed Central (United Kingdom)

The common marmoset (Callithrix jacchus) is increasingly attractive for use as a non-human primate animal model in biomedical research. It has a relatively high reproduction rate for a primate, making it potentially suitable for transgenic modification. Although several attempts have been made to produce non-human transgenic primates, transgene expression in the somatic tissues of live infants has not been demonstrated by objective analyses such as polymerase chain reaction with reverse transcription or western blots. Here we show that the injection of a self-inactivating lentiviral vector in sucrose solution into marmoset embryos results in transgenic common marmosets that expressed the transgene in several organs. Notably, we achieved germline transmission of the transgene, and the transgenic offspring developed normally. The successful creation of transgenic marmosets provides a new animal model for human disease that has the great advantage of a close genetic relationship with humans. This model will be valuable to many fields of biomedical research.

Sasaki E; Suemizu H; Shimada A; Hanazawa K; Oiwa R; Kamioka M; Tomioka I; Sotomaru Y; Hirakawa R; Eto T; Shiozawa S; Maeda T; Ito M; Ito R; Kito C; Yagihashi C; Kawai K; Miyoshi H; Tanioka Y; Tamaoki N; Habu S; Okano H; Nomura T

2009-05-01

 
 
 
 
281

Lectin cDNA and transgenic plants derived therefrom  

Energy Technology Data Exchange (ETDEWEB)

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

Raikhel, Natasha V. (Okemos, MI)

2000-10-03

282

Insect-resistant transgenic brinjal plants.  

UK PubMed Central (United Kingdom)

A synthetic cry1Ab gene coding for an insecticidal crystal protein (ICP) of Bacillus thuringiensis (Bt) was transferred to brinjal (eggplant) by cocultivating cotyledonary explants with Agrobacterium tumefaciens. Transformant plants resistant to kanamycin were regenerated. Hybridization experiments demonstrated gene integration and mRNA expression. Double-antibody sandwich ELISA analysis revealed Bt toxin protein expression in the transgenic plants. The expression resulted in a significant insecticidal activity of transgenic brinjal fruits against the larvae of fruit borer (Leucinodes orbonalis). The results also demonstrated that a synthetic gene based on monocot codon usage can be expressed in dicotyledonous plants for insect control.

Kumar PA; Mandaokar A; Sreenivasu K; Chakrabarti SK; Bisaria S; Sharma SR; Kaur S; Sharma RP

1998-01-01

283

Edible Transgenic Plant Vaccines for Different Diseases.  

UK PubMed Central (United Kingdom)

Edible plant vaccines are immunogenic preparations containing antigenic proteins rather than pathogens, therefore, they sanctify situation where there is a possibility of resurgence of disease when the antigenic preparation contains the organism in any form whatsoever. Expression of antigens as vaccines and of antibodies against antigens of pathogens in transgenic plants is a convenient and inexpensive source for various bacterial, viral, helminths, protozoan and autoimmune diseases with lower capital costs. This review describes various diseases along with the production of edible transgenic plant vaccines/ proteins for the same. Thus, substituting and improvising conventional immunization methods.

Jain A; Saini V; Veer Kohli D

2013-09-01

284

Genomic stability and long-term transgene expression in poplar.  

Science.gov (United States)

Stable expression of foreign genes over the entire life span of a plant is important for long-lived organisms such as trees. For transgenic forest trees, very little information is available on long-term transgene expression and genomic stability. Independent transgenic lines obtained directly after transformation are initially screened in respect to T-DNA integration and transgene expression. However, very little consideration has been given to long-term transgene stability in long-lived forest trees. We have investigated possible genome wide changes following T-DNA integration as well as long-term stability of transgene expression in different transgenic lines of hybrid aspen (Populus tremula × Populus tremuloides) that are up to 19 years old. For studies on possible genome wide changes following T-DNA integration, four different independent rolC-transgenic lines were subjected to an extensive AFLP study and compared to the non-transgenic control line. Only minor genomic changes following T-DNA integration could be detected. To study long-term transgene expression, six different independent rolC-transgenic lines produced in 1993 and since that time have been kept continuously under in vitro conditions. In addition, 18 transgenic plants belonging to eight independent rolC-transgenic lines transferred to glasshouse between 1994 and 2004 were chosen to determine the presence and expression of the rolC gene. In all transgenic lines examined, the rolC gene could successfully be amplified by PCR tests. Both, the 19 years old tissue cultures and the up to 18 years old glasshouse-grown trees revealed expression of the rolC transgene, as demonstrated by the rolC-phenotype and/or northern blot experiments confirming long-term transgene expression. PMID:23740206

Fladung, Matthias; Hoenicka, Hans; Raj Ahuja, M

2013-06-01

285

Genomic stability and long-term transgene expression in poplar.  

UK PubMed Central (United Kingdom)

Stable expression of foreign genes over the entire life span of a plant is important for long-lived organisms such as trees. For transgenic forest trees, very little information is available on long-term transgene expression and genomic stability. Independent transgenic lines obtained directly after transformation are initially screened in respect to T-DNA integration and transgene expression. However, very little consideration has been given to long-term transgene stability in long-lived forest trees. We have investigated possible genome wide changes following T-DNA integration as well as long-term stability of transgene expression in different transgenic lines of hybrid aspen (Populus tremula × Populus tremuloides) that are up to 19 years old. For studies on possible genome wide changes following T-DNA integration, four different independent rolC-transgenic lines were subjected to an extensive AFLP study and compared to the non-transgenic control line. Only minor genomic changes following T-DNA integration could be detected. To study long-term transgene expression, six different independent rolC-transgenic lines produced in 1993 and since that time have been kept continuously under in vitro conditions. In addition, 18 transgenic plants belonging to eight independent rolC-transgenic lines transferred to glasshouse between 1994 and 2004 were chosen to determine the presence and expression of the rolC gene. In all transgenic lines examined, the rolC gene could successfully be amplified by PCR tests. Both, the 19 years old tissue cultures and the up to 18 years old glasshouse-grown trees revealed expression of the rolC transgene, as demonstrated by the rolC-phenotype and/or northern blot experiments confirming long-term transgene expression.

Fladung M; Hoenicka H; Raj Ahuja M

2013-06-01

286

Activation of polyamine catabolism in transgenic rats induces acute pancreatitis  

Digital Repository Infrastructure Vision for European Research (DRIVER)

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 ...

Alhonen, Leena; Parkkinen, Jyrki J.; Keinänen, Tuomo; Sinervirta, Riitta; Herzig, Karl-Heinz; Jänne, Juhani

287

Growth and endocrine effects of recombinant bovine growth hormone treatment in non-transgenic and growth hormone transgenic coho salmon.  

UK PubMed Central (United Kingdom)

To examine the relative growth, endocrine, and gene expression effects of growth hormone (GH) transgenesis vs. GH protein treatment, wild-type non-transgenic and GH transgenic coho salmon were treated with a sustained-release formulation of recombinant bovine GH (bGH; Posilac). Fish size, specific growth rate (SGR), and condition factor (CF) were monitored for 14 weeks, after which endocrine parameters were measured. Transgenic fish had much higher growth, SGR and CF than non-transgenic fish, and bGH injection significantly increased weight and SGR in non-transgenic but not transgenic fish. Plasma salmon GH concentrations decreased with bGH treatment in non-transgenic but not in transgenic fish where levels were similar to controls. Higher GH mRNA levels were detected in transgenic muscle and liver but no differences were observed in GH receptor (GHR) mRNA levels. In non-transgenic pituitary, GH and GHR mRNA levels per mg pituitary decreased with bGH dose to levels seen in transgenic salmon. Plasma IGF-I was elevated with bGH dose only in non-transgenic fish, while transgenic fish maintained an elevated level of IGF-I with or without bGH treatment. A similar trend was seen for liver IGF-I mRNA levels. Thus, bGH treatment increased fish growth and influenced feedback on endocrine parameters in non-transgenic but not in transgenic fish. A lack of further growth stimulation of GH transgenic fish suggests that these fish are experiencing maximal growth stimulation via GH pathways.

Raven PA; Sakhrani D; Beckman B; Neregård L; Sundström LF; Björnsson BT; Devlin RH

2012-05-01

288

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

Directory of Open Access Journals (Sweden)

Full Text Available Among all the transgenic mammalians produced so far, goats have represented an excellent model of transgenesis when considering the factors such as the market demand for protein, volume of milk produced per lactation and reproductive rate. Various recombinant proteins have been obtained from the transgenic and non-transgenic goats, and among these, human antithrombin, produced by the transgenic goats, was the first recombinant protein of animal origin to be released as a drug for the clinical use in humans. This review reports the aspects inherent to the production of recombinant proteins in the goats, from the production of the animal bioreactors up to the expression of these proteins in their milk.

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

2011-01-01

289

Comparative analysis of nutritional compositions of transgenic high iron rice with its non-transgenic counterpart.  

UK PubMed Central (United Kingdom)

Iron is an essential micronutrient for human nutrition and polished rice contains very low amount of iron. Rice with high iron content in seed endosperm has been developed by insertion of soybean ferritin gene under the control of the endosperm specific glutelin promoter into the genome of indica rice line IR68144. The nutritional composition of the brown and milled rice grain has been compared with that of the non-transgenic rice of the same variety. In this study, the nutritional components, as well as the anti-nutrient levels, were measured. Our studies established that apart from the increased level of iron and zinc in transgenic seeds, the nutritional quality of both the brown and milled rice grains from the transgenic line was substantially equivalent to that of the non-transgenic rice plants. The result clearly shows that the measured amounts of the nutritional components are well within the range of values reported for other commercial lines.

Gayen D; Sarkar SN; Datta SK; Datta K

2013-06-01

290

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

291

Non-transgenic herbicide resistant plants  

UK PubMed Central (United Kingdom)

The present invention relates to the production of a non-transgenic plant resistant or tolerant to a herbicide of the phosphonomethylglycinc family, e.g., glyphosate. The present invention also relates to the use of a recombinagenic Oligonucleobase to make a desired mutation in the chromosomal or episomal sequences of a plant in the gene encoding for 5-enol pyruvyl-shikimate-3-phosphate synthase (EPSPS). The mutated protein, which substantially maintains the catalytic activity of the wild-type protein, allows for increased resistance or tolerance of the plant to a herbicide of the phosphonomethylglycine family, and allows for the substantially normal growth or development of the plant, its organs, tissues or cells as compared to the wild-type plant irrespective of the presence or absence of the herbicide. The present invention also relates to a non-transgenic plant cell in which the EPSPS gene has been mutated, a non-transgenic plant regenerated therefrom, as well as a plant resulting from a cross using a regenerated non-transgenic plant having a mutated EPSPS gene.

BEETHAM PETER R; AVISSAR PATRICIA L; WALKER KEITH A; METZ RICHARD A

292

Non-transgenic herbicide resistant plants  

UK PubMed Central (United Kingdom)

The present invention relates to the production of a non-transgenic plant resistant or tolerant to a herbicide of the phosphonomethylglycine family, e.g., glyphosate. The present invention also relates to the use of a recombinagenic Oligonucleobase to make a desired mutation in the chromosomal or episomal sequences of a plant in the gene encoding for 5-enol pyruvylshikimate-3-phosphate synthase (EPSPS). The mutated protein, which substantially maintains the catalytic activity of the wild-type protein, allows for increased resistance or tolerance of the plant to a herbicide of the phosphonomethylglycine family, and allows for the substantially normal growth or development of the plant, its organs, tissues or cells as compared to the wild-type plant irrespective of the presence or absence of the herbicide. The present invention also relates to a non-transgenic plant cell in which the EPSPS gene has been mutated, a non-transgenic plant regenerated therefrom, as well as a plant resulting from a cross using a regenerated non-transgenic plant having a mutated EPSPS gene.

BEETHAM PETER R; AVISSAR PATRICIA L; WALKER KEITH A; METZ RICHARD A

293

Metal resistance sequences and transgenic plants  

Energy Technology Data Exchange (ETDEWEB)

The present invention provides nucleic acid sequences encoding a metal ion resistance protein, which are expressible in plant cells. The metal resistance protein provides for the enzymatic reduction of metal ions including but not limited to divalent Cu, divalent mercury, trivalent gold, divalent cadmium, lead ions and monovalent silver ions. Transgenic plants which express these coding sequences exhibit increased resistance to metal ions in the environment as compared with plants which have not been so genetically modified. Transgenic plants with improved resistance to organometals including alkylmercury compounds, among others, are provided by the further inclusion of plant-expressible organometal lyase coding sequences, as specifically exemplified by the plant-expressible merB coding sequence. Furthermore, these transgenic plants which have been genetically modified to express the metal resistance coding sequences of the present invention can participate in the bioremediation of metal contamination via the enzymatic reduction of metal ions. Transgenic plants resistant to organometals can further mediate remediation of organic metal compounds, for example, alkylmetal compounds including but not limited to methyl mercury, methyl lead compounds, methyl cadmium and methyl arsenic compounds, in the environment by causing the freeing of mercuric or other metal ions and the reduction of the ionic mercury or other metal ions to the less toxic elemental mercury or other metals.

Meagher, Richard Brian (Athens, GA); Summers, Anne O. (Athens, GA); Rugh, Clayton L. (Athens, GA)

1999-10-12

294

Transgenic Mouse Model of Chronic Beryllium Disease  

Energy Technology Data Exchange (ETDEWEB)

Animal models provide powerful tools for dissecting dose-response relationships and pathogenic mechanisms and for testing new treatment paradigms. Mechanistic research on beryllium exposure-disease relationships is severely limited by a general inability to develop a sufficient chronic beryllium disease animal model. Discovery of the Human Leukocyte Antigen (HLA) - DPB1Glu69 genetic susceptibility component of chronic beryllium disease permitted the addition of this human beryllium antigen presentation molecule to an animal genome which may permit development of a better animal model for chronic beryllium disease. Using FVB/N inbred mice, Drs. Rubin and Zhu, successfully produced three strains of HLA-DPB1 Glu 69 transgenic mice. Each mouse strain contains a haplotype of the HLA-DPB1 Glu 69 gene that confers a different magnitude of odds ratio (OR) of risk for chronic beryllium disease: HLA-DPB1*0401 (OR = 0.2), HLA-DPB1*0201 (OR = 15), HLA-DPB1*1701 (OR = 240). In addition, Drs. Rubin and Zhu developed transgenic mice with the human CD4 gene to permit better transmission of signals between T cells and antigen presenting cells. This project has maintained the colonies of these transgenic mice and tested the functionality of the human transgenes.

Gordon, Terry

2009-05-26

295

Transgenic plants containing soluble cell wall polysaccharides  

UK PubMed Central (United Kingdom)

The present invention relates to a transgenic plant expressing CDH. The invention furthermore relates to s method of making a bio-fuel, wood, paper, textile or yarn product, comprising: a) over-expressing a nucleic acid molecule encoding CDH in a plant and b) processing the cellulose or hemicellulose portion of the plant into said product.

SHANI ZIV; SHOSEYOV ODED; ABRAMSON MIRON; BARIMBOIM NOGA; DEKEL MARA; LAPIDOT SHAUL

296

Transgenic trees: how far the pollen flies?  

Directory of Open Access Journals (Sweden)

Full Text Available Implications of extended gene flow is discussed under the light of new laws issued, regulating the admixtures among organic, traditional and biotec agriculture. Recent scientific literature is reviewed on the role of gene flow in gene escape in transgenic forest trees.

Leonardi S

2006-01-01

297

Ethical Issues in Genetic Engineering and Transgenics  

Science.gov (United States)

The issue-focused, peer-reviewed article demonstrates how transgenic technology has the potential of medical therapy, but it raises questions about these issues: creation of new life forms and crossing species boundaries, long-term effects on human health and the environment, blending of nonhuman animal and human DNA , and unintended personal, social, and cultural consequences.

Linda MacDonald Glenn (;)

2004-06-01

298

Analysis of somatic mutations in kappa transgenes  

Digital Repository Infrastructure Vision for European Research (DRIVER)

We have examined the nature and localization of somatic mutations in three kappa transgenes cloned from IgG-secreting hybridomas. All of the mutations identified were single base substitutions. Mutations were localized to the variable (V) region and its flanking sequences. In every case, the nuclear...

299

Can Transgenic Maize Affect Soil Microbial Communities?  

Digital Repository Infrastructure Vision for European Research (DRIVER)

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 gu...

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

300

Monitoring transgenic plants using in vivo markers  

Energy Technology Data Exchange (ETDEWEB)

The gene coding for green fluorecent protein (GFP), isolated and cloned from the jellyfish Aequorea victoria, is an ideal transgene for the monitoring of any plant species. It has the ability to fluoresce without added substrate, enzyme, or cofactor; it does not introduce morphological or sexual aberrations when expressed. 7 refs., 1 fig.

Stewart, C.N. Jr. [Univ. of North Carolina, Greensboro, NC (United States)

1996-06-01

 
 
 
 
301

Progress in Xenotransplantation Research Employing Transgenic Pigs  

Directory of Open Access Journals (Sweden)

Full Text Available Microinjection of foreign DNA into pronuclei of a fertilized oocyte has predominantly been used for the generation of transgenic livestock. This technology works reliably, but is inefficient and results in random integration and variable expression patterns in the transgenic offspring. Nevertheless, remarkable achievements have been made with this technology with regard to xenotransplantation. Transgenic pigs that express human complement regulating proteins have been tested in their ability to serve as donors in human organ transplantation (i.e. xenotransplantation). In vitro and in vivo data convincingly show that the hyperacute rejection response can be overcome in a clinically acceptable manner by successfully employing this strategy. The recent developments in nuclear transfer and its merger with the growing genomic data allow targeted and regulatable transgenesis. Systems for efficient homologous recombination in somatic cells are being developed and the first knock-out pigs, carrying a deletion in the a-galactosyltransferase gene, were recently generated. It is anticipated that poly-transgenic pigs will be available as donors for functional xenografts within a few years. Similarly, pigs may serve as donors for a variety of xenogenic cells and tissues. The availability of these technologies is essential to maintain "genetic security" and to ensure absence of unwanted side effects.

H. Niemann; W. A. Kues

2003-01-01

302

A triple-transgenic immunotolerant mouse model.  

UK PubMed Central (United Kingdom)

Avoiding unwanted immunogenicity is of key importance in the development of therapeutic drug proteins. Animal models are of less predictive value because most of the drug proteins are recognized as foreign proteins. However, different methods have been developed to obtain immunotolerant animal models. So far, the immunotolerant animal models have been developed to assess one protein at a time and are not suitable for the assessment of combination products. Our aim was to develop an animal model for evaluating the impact of manufacturing and formulation changes on immunogenicity, suitable for both single protein and combination products. We constructed two lines of transgenic mice expressing the three human coagulation factors, II, VII, and X, by inserting a single vector containing the three coagulation factors encoding sequences separated by insulator sequences derived from the chicken beta-globin locus into the mouse genome. Immunization of transgenic mice from the two lines and their wild-type littermates showed that transgenic mice from both lines were immunotolerant to the expressed human coagulation factors. We conclude that transgenic mice immunotolerant to multiple proteins can be obtained, and that these mice are potentially useful as animal models in the assessment of immunogenicity in response to manufacturing changes.

Brenden N; Madeyski-Bengtson K; Martinsson K; Svärd R; Albery-Larsdotter S; Granath B; Lundgren H; Lövgren A

2013-03-01

303

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.

1994-01-01

304

Approaching the Lower Limits of Transgene Variability.  

Science.gov (United States)

The inclusion of chicken lysozyme matrix-associated regions (MARs) in T-DNA has been demonstrated to reduce the variation in [beta]-glucuronidase (GUS) gene expression among first-generation transformed plants. The residual variation observed between transgenic plant lines with MARs at the T-DNA borders was investigated. By definition, any phenotypic variance between or within genetically identical plants is caused by random or environmental variation. This variation therefore sets a lower limit to the variation in GUS activities. The variance of GUS activity in offspring plant populations of genetically identical individuals was used as an estimate of environmental variation. For transgenic plants with MARs at the T-DNA borders, the variation between independent transformants could not be distinguished from the environmental variation. The variation could be attributed mainly to the variation in the GUS activity measurement. Therefore, the MAR element approached the maximal possible reduction of transgene variability given current technology and sample sizes. The role of MARs in offspring plants was evaluated by comparing such populations of transgenic plants for the magnitude of and variation in GUS activity. Pairwise comparisons showed that the presence of MARs reduced variation in offspring generations in the same manner as demonstrated for primary transformants. The populations carrying a doubled cauliflower mosaic virus 35S promoter-GUS gene tended to be more variable than the Lhca3.St.1 promoter-GUS gene-carrying populations. This tendency indicated an intrinsic susceptibility of the doubled cauliflower mosaic virus 35S promoter to variation. Homozygous plants were approximately twice as active as the corresponding hemizygous plants and tended to be more variable than the hemizygous plants. We hypothesized that the magnitude of environmental variations is related to a higher susceptibility to transgene silencing. PMID:12239419

Mlynarova, L.; Keizer, LCP.; Stiekema, W. J.; Nap, J. P.

1996-09-01

305

Approaching the Lower Limits of Transgene Variability.  

UK PubMed Central (United Kingdom)

The inclusion of chicken lysozyme matrix-associated regions (MARs) in T-DNA has been demonstrated to reduce the variation in [beta]-glucuronidase (GUS) gene expression among first-generation transformed plants. The residual variation observed between transgenic plant lines with MARs at the T-DNA borders was investigated. By definition, any phenotypic variance between or within genetically identical plants is caused by random or environmental variation. This variation therefore sets a lower limit to the variation in GUS activities. The variance of GUS activity in offspring plant populations of genetically identical individuals was used as an estimate of environmental variation. For transgenic plants with MARs at the T-DNA borders, the variation between independent transformants could not be distinguished from the environmental variation. The variation could be attributed mainly to the variation in the GUS activity measurement. Therefore, the MAR element approached the maximal possible reduction of transgene variability given current technology and sample sizes. The role of MARs in offspring plants was evaluated by comparing such populations of transgenic plants for the magnitude of and variation in GUS activity. Pairwise comparisons showed that the presence of MARs reduced variation in offspring generations in the same manner as demonstrated for primary transformants. The populations carrying a doubled cauliflower mosaic virus 35S promoter-GUS gene tended to be more variable than the Lhca3.St.1 promoter-GUS gene-carrying populations. This tendency indicated an intrinsic susceptibility of the doubled cauliflower mosaic virus 35S promoter to variation. Homozygous plants were approximately twice as active as the corresponding hemizygous plants and tended to be more variable than the hemizygous plants. We hypothesized that the magnitude of environmental variations is related to a higher susceptibility to transgene silencing.

Mlynarova L; Keizer L; Stiekema WJ; Nap JP

1996-09-01

306

Transgenic pig carrying green fluorescent proteasomes  

Science.gov (United States)

Among its many functions, the ubiquitin–proteasome system regulates substrate-specific proteolysis during the cell cycle, apoptosis, and fertilization and in pathologies such as Alzheimer’s disease, cancer, and liver cirrhosis. Proteasomes are present in human and boar spermatozoa, but little is known about the interactions of proteasomal subunits with other sperm proteins or structures. We have created a transgenic boar with green fluorescent protein (GFP) tagged 20S proteasomal core subunit ?-type 1 (PSMA1-GFP), hypothesizing that the PSMA1-GFP fusion protein will be incorporated into functional sperm proteasomes. Using direct epifluorescence imaging and indirect immunofluorescence detection, we have confirmed the presence of PSMA1-GFP in the sperm acrosome. Western blotting revealed a protein band corresponding to the predicted mass of PSMA1-GFP fusion protein (57 kDa) in transgenic spermatozoa. Transgenic boar fertility was confirmed by in vitro fertilization, resulting in transgenic blastocysts, and by mating, resulting in healthy transgenic offspring. Immunoprecipitation and proteomic analysis revealed that PSMA1-GFP copurifies with several acrosomal membrane-associated proteins (e.g., lactadherin/milk fat globule E8 and spermadhesin alanine-tryptophan-asparagine). The interaction of MFGE8 with PSMA1-GFP was confirmed through cross-immunoprecipitation. The identified proteasome-interacting proteins may regulate sperm proteasomal activity during fertilization or may be the substrates of proteasomal proteolysis during fertilization. Proteomic analysis also confirmed the interaction/coimmunoprecipitation of PSMA1-GFP with 13/14 proteasomal core subunits. These results demonstrate that the PSMA1-GFP was incorporated in the assembled sperm proteasomes. This mammal carrying green fluorescent proteasomes will be useful for studies of fertilization and wherever the ubiquitin–proteasome system plays a role in cellular function or pathology.

Miles, Edward L.; O'Gorman, Chad; Zhao, Jianguo; Samuel, Melissa; Walters, Eric; Yi, Young-Joo; Prather, Randall S.; Wells, Kevin D.; Sutovsky, Peter

2013-01-01

307

Transgenic pig carrying green fluorescent proteasomes.  

UK PubMed Central (United Kingdom)

Among its many functions, the ubiquitin-proteasome system regulates substrate-specific proteolysis during the cell cycle, apoptosis, and fertilization and in pathologies such as Alzheimer's disease, cancer, and liver cirrhosis. Proteasomes are present in human and boar spermatozoa, but little is known about the interactions of proteasomal subunits with other sperm proteins or structures. We have created a transgenic boar with green fluorescent protein (GFP) tagged 20S proteasomal core subunit ?-type 1 (PSMA1-GFP), hypothesizing that the PSMA1-GFP fusion protein will be incorporated into functional sperm proteasomes. Using direct epifluorescence imaging and indirect immunofluorescence detection, we have confirmed the presence of PSMA1-GFP in the sperm acrosome. Western blotting revealed a protein band corresponding to the predicted mass of PSMA1-GFP fusion protein (57 kDa) in transgenic spermatozoa. Transgenic boar fertility was confirmed by in vitro fertilization, resulting in transgenic blastocysts, and by mating, resulting in healthy transgenic offspring. Immunoprecipitation and proteomic analysis revealed that PSMA1-GFP copurifies with several acrosomal membrane-associated proteins (e.g., lactadherin/milk fat globule E8 and spermadhesin alanine-tryptophan-asparagine). The interaction of MFGE8 with PSMA1-GFP was confirmed through cross-immunoprecipitation. The identified proteasome-interacting proteins may regulate sperm proteasomal activity during fertilization or may be the substrates of proteasomal proteolysis during fertilization. Proteomic analysis also confirmed the interaction/coimmunoprecipitation of PSMA1-GFP with 13/14 proteasomal core subunits. These results demonstrate that the PSMA1-GFP was incorporated in the assembled sperm proteasomes. This mammal carrying green fluorescent proteasomes will be useful for studies of fertilization and wherever the ubiquitin-proteasome system plays a role in cellular function or pathology.

Miles EL; O'Gorman C; Zhao J; Samuel M; Walters E; Yi YJ; Sutovsky M; Prather RS; Wells KD; Sutovsky P

2013-04-01

308

Commercialization of transgenic rice in China: potential environmental biosafety issues  

Directory of Open Access Journals (Sweden)

Full Text Available The development and commercialization of transgenic rice with novel traits in China may offer more opportunities for promoting rice productivity. Owing to the significance of rice as a major food crop in China, the enhancement of rice production is important for national food security. If left unaddressed, the potential biosafety concerns over the extensive release and commercial cultivation of transgenic rice may hamper the development and application of this technology in rice improvement. Biosafety issues include: (1) effectsof toxic transgenes on non-target organisms; (2) transgene escape to crops or wild relatives through gene flow and its potential ecological consequences; (3) interactions and influences of transgenes and transgenic plants on biodiversity, ecosystem functions, and soil microbes; and (4) the development of resistance to insect- or disease-resistant transgenes in target organisms. In order to safely and sustainably utilize transgenic biotechnology in rice, it is very important to assess biosafety consequences, including environmental risks, from transgenic rice. This paper presents a rational analysis of potential environmental biosafety problems based on the principles of risk assessment, provided that transgenic rice will be released for commercialization. We hope these analyses will provide useful information for the decision-making on commercialization of transgenic rice and serve as a framework for the assessment of relevant environmental biosafety risks.

Bao-Rong Lu; Qiang Fu; Zhicheng Shen

2008-01-01

309

Development of expression vectors for transgenic fish.  

Science.gov (United States)

Genetic alteration of fish is important for aquatic biotechnology as well as for investigating molecular interactions that occur during vertebrate development. The numerous, large, transparent, and externally fertilized eggs of many fish species make them ideally suitable for genetic manipulation, especially for production of transgenic animals. Genetic engineering of fish requires suitable expression vectors. Accordingly, we developed two fish expression vectors, FV-1 and FV-2, which contain the proximal promoter and enhancer regulatory elements of the carp beta-actin gene and the polyadenylation signal from the salmon growth hormone gene. The two fish expression vectors were tested in microinjected fish eggs and in tissue cultured fish and mammalian cells. These two "all-fish" expression vectors should be useful for genetic engineering of fish and have been used with growth-enhancing genes in transgenic fish. PMID:1366961

Liu, Z J; Moav, B; Faras, A J; Guise, K S; Kapuscinski, A R; Hackett, P B

1990-12-01

310

A multidisciplinary approach involving comparative 'OMICS' of transgenic and non-transgenic soybeam seeds  

International Nuclear Information System (INIS)

[en] Complete text of publication follows. Soybean culture has an expressive impact in the economy of many countries, being the commercialization of its by-products, which presents many benefits in terms of health and nutritional aspects, and which also includes a fuel alternative (biodiesel), the main factor for the large soybean production. Part of this impact is due to the transgenic modification of soybean, conferring enhanced characteristics to the culture, such as tolerance to fungicides (Y. Kim et al., J. Microbiol. Biotechnol., 16 (2006), 25-31.). Due to the insertion of hexogen genes, some proteome modification is possible (S. Natarajan et al., Anal. Biochem., 342 (2005), 214-220), and recently some metallome modification was reported by our research group (A. Sussulini et al., J. Anal. At. Spectrom., 22 (2007), 1501-1506). Then, the aim of this work is to enlarge the results in terms of 'omics' when considering transgenic and non-transgenic soybean seeds. For this task, the identification of more than 140 soybean proteins using MALDI-QTOF-MS after 2D-PAGE protein separation (369±46 and 376±42 protein spots in the 4-7 pI range for transgenic and non-transgenic soybean seeds, respectively), the analysis of the protein expression using image program, the analysis of some enzymes (SOD, GR, APX, CAT) involved in the ROS production, the mapping of 80 protein spots using SR-XRF, and the metal identification of more than 30 spots using ICP-MS was carried out. In terms of metal distribution when considering some proteins, the results displayed a great ability of proteins bind different metal ions. High iron (sucrose binding protein homolog S-64 - 57,922 kDa), chromium (protein not identified), lead (seed maturation protein PM 41 - 15,103 kDa), copper and tin (trypsin inhibitor (kunitz), chain A - 20,417 kDa) contents were achieved in the non-transgenic soybean, while high magnesium (actin - 50,281 kDa), barium (protein not identified) and ruthenium (protein not identified) contents were achieved in the transgenic soybean. The results put in evidence the possibility to find a biomarker candidate for differentiating transgenic and non-transgenic organisms. Financial support from Fundacao de Amparo a Pesquisa do Estado de Sao Paulo - FAPESP, Conselho Nacional de Desenvolvimento Cientifico e Tecnologico - CNPq, Fianciadora de Estudos e Projetos - FINEP, and Proteome Network of the Sao Paulo state - Brazilian National Laboratory of Synchrotron Radiation are highly acknowledged.

2009-09-03

311

Green tea polyphenols control dysregulated glutamate dehydrogenase in transgenic mice by hijacking the ADP activation site.  

Science.gov (United States)

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 ?-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. PMID:21813650

Li, Changhong; Li, Ming; Chen, Pan; Narayan, Srinivas; Matschinsky, Franz M; Bennett, Michael J; Stanley, Charles A; Smith, Thomas J

2011-08-03

312

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

UK PubMed Central (United Kingdom)

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.

Haroldsen VM; Chi-Ham CL; Bennett AB

2012-10-01

313

A Transgenic Tri-Modality Reporter Mouse  

Science.gov (United States)

Transgenic mouse with a stably integrated reporter gene(s) can be a valuable resource for obtaining uniformly labeled stem cells, tissues, and organs for various applications. We have generated a transgenic mouse model that ubiquitously expresses a tri-fusion reporter gene (fluc2-tdTomato-ttk) driven by a constitutive chicken ?-actin promoter. This “Tri-Modality Reporter Mouse” system allows one to isolate most cells from this donor mouse and image them for bioluminescent (fluc2), fluorescent (tdTomato), and positron emission tomography (PET) (ttk) modalities. Transgenic colonies with different levels of tri-fusion reporter gene expression showed a linear correlation between all three-reporter proteins (R2=0.89 for TdTomato vs Fluc, R2=0.94 for Fluc vs TTK, R2=0.89 for TdTomato vs TTK) in vitro from tissue lysates and in vivo by optical and PET imaging. Mesenchymal stem cells (MSCs) isolated from this transgenics showed high level of reporter gene expression, which linearly correlated with the cell numbers (R2=0.99 for bioluminescence imaging (BLI)). Both BLI (R2=0.93) and micro-PET (R2=0.94) imaging of the subcutaneous implants of Tri-Modality Reporter Mouse derived MSCs in nude mice showed linear correlation with the cell numbers and across different imaging modalities (R2=0.97). Serial imaging of MSCs transplanted to mice with acute myocardial infarction (MI) by intramyocardial injection exhibited significantly higher signals in MI heart at days 2, 3, 4, and 7 (pstem cell research, tissue engineering research, and organ transplantation.

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

2013-01-01

314

Regulation of transgene expression in genetic immunization  

Directory of Open Access Journals (Sweden)

Full Text Available The use of mammalian gene expression vectors has become increasingly important for genetic immunization and gene therapy as well as basic research. Essential for the success of these vectors in genetic immunization is the proper choice of a promoter linked to the antigen of interest. Many genetic immunization vectors use promoter elements from pathogenic viruses including SV40 and CMV. Lymphokines produced by the immune response to proteins expressed by these vectors could inhibit further transcription initiation by viral promoters. Our objective was to determine the effect of IFN-g on transgene expression driven by viral SV40 or CMV promoter/enhancer and the mammalian promoter/enhancer for the major histocompatibility complex class I (MHC I) gene. We transfected the luciferase gene driven by these three promoters into 14 cell lines of many tissues and several species. Luciferase assays of transfected cells untreated or treated with IFN-g indicated that although the viral promoters could drive luciferase production in all cell lines tested to higher or lower levels than the MHC I promoter, treatment with IFN-g inhibited transgene expression in most of the cell lines and amplification of the MHC I promoter-driven transgene expression in all cell lines. These data indicate that the SV40 and CMV promoter/enhancers may not be a suitable choice for gene delivery especially for genetic immunization or cancer cytokine gene therapy. The MHC I promoter/enhancer, on the other hand, may be an ideal transgene promoter for applications involving the immune system.

J.S. Harms; S.C. Oliveira; G.A. Splitter

1999-01-01

315

Regulation of transgene expression in genetic immunization  

Scientific Electronic Library Online (English)

Full Text Available Abstract in english The use of mammalian gene expression vectors has become increasingly important for genetic immunization and gene therapy as well as basic research. Essential for the success of these vectors in genetic immunization is the proper choice of a promoter linked to the antigen of interest. Many genetic immunization vectors use promoter elements from pathogenic viruses including SV40 and CMV. Lymphokines produced by the immune response to proteins expressed by these vectors coul (more) d 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.; Oliveira, S.C.; Splitter, G.A.

1999-02-01

316

Transgenic analysis of GFAP promoter elements.  

UK PubMed Central (United Kingdom)

Transcriptional regulation of the glial fibrillary acidic protein gene (GFAP) is of interest because of its astrocyte specificity and its upregulation in response to CNS injuries. We have used a transgenic approach instead of cell transfection to identify promoter elements of the human GFAP gene, since previous observations show that GFAP transcription is regulated differently in transfected cultured cells from in the mouse. We previously showed that block mutation of enhancer regions spanning from bp -1488 to -1434 (the C1.1 segment) and -1443 to -1399 (C1.2) resulted in altered patterns of expression and loss of astrocyte specificity, respectively. This analysis has now been extended upstream to bp -1612 to -1489 (the B region), which previously has been shown especially important for expression. Block mutation of each of four contiguous sequences, which together span the B region, each decreased the level of transgene activity by at least 50%, indicating that multiple sites contribute to the transcriptional activity in a cooperative manner. Several of the block mutations also altered the brain region pattern of expression, astrocyte specificity and/or the developmental time course. Transgenes were then analyzed in which mutations were limited to specific transcription factor binding sites in each of the 4 B block segments as well as in C1.1 and C1.2. Whereas mutation of the conserved consensus AP-1 site unexpectedly had little effect on transgene expression; NFI, SP1, STAT3, and NF-?B were identified as having important roles in regulating the strength of GFAP promoter activity and/or its astrocyte specificity.

Yeo S; Bandyopadhyay S; Messing A; Brenner M

2013-09-01

317

Transgenic analysis of GFAP promoter elements.  

Science.gov (United States)

Transcriptional regulation of the glial fibrillary acidic protein gene (GFAP) is of interest because of its astrocyte specificity and its upregulation in response to CNS injuries. We have used a transgenic approach instead of cell transfection to identify promoter elements of the human GFAP gene, since previous observations show that GFAP transcription is regulated differently in transfected cultured cells from in the mouse. We previously showed that block mutation of enhancer regions spanning from bp -1488 to -1434 (the C1.1 segment) and -1443 to -1399 (C1.2) resulted in altered patterns of expression and loss of astrocyte specificity, respectively. This analysis has now been extended upstream to bp -1612 to -1489 (the B region), which previously has been shown especially important for expression. Block mutation of each of four contiguous sequences, which together span the B region, each decreased the level of transgene activity by at least 50%, indicating that multiple sites contribute to the transcriptional activity in a cooperative manner. Several of the block mutations also altered the brain region pattern of expression, astrocyte specificity and/or the developmental time course. Transgenes were then analyzed in which mutations were limited to specific transcription factor binding sites in each of the 4 B block segments as well as in C1.1 and C1.2. Whereas mutation of the conserved consensus AP-1 site unexpectedly had little effect on transgene expression; NFI, SP1, STAT3, and NF-?B were identified as having important roles in regulating the strength of GFAP promoter activity and/or its astrocyte specificity. PMID:23832770

Yeo, Sujeong; Bandyopadhyay, Susanta; Messing, Albee; Brenner, Michael

2013-07-08

318

Transgene overexpression of alphaB crystallin confers simultaneous protection against cardiomyocyte apoptosis and necrosis during myocardial ischemia and reperfusion.  

Science.gov (United States)

We investigated whether enhanced expression of alphaB crystallin, a stress-inducible molecular chaperone of the small heat shock family, can protect myocardial contractile apparatus against ischemia reperfusion (I/R) injury. Transgenic mice overexpressing alphaB crystallin were generated using the 0.76 kb rat alphaB crystallin cDNA cloned into a pCAGGS plasmid driven by a human cytomegalovirus expression system. Southern analysis confirmed transgene integration and Northern and Western blotting characterized expression (3.1-fold and 6.9-fold elevations in myocardial mRNA and protein levels, respectively). Extent of functional recovery over a 3 h reperfusion period following a 20 min ischemic period in transgenic and wild-type mouse hearts was assessed using an ex vivo work-performing heart preparation. The transgenic group displayed significantly higher values of DP at R45 min (29.14+/-1.9 mm Hg vs. 17.6+/-0.7 mm Hg), R60 min (31.56+/-1.7 mm Hg vs. 17.8+/-0.8 mm Hg), and R75 min (32.5+/-2.2 mm Hg vs. 16.9+/-0.9 mm Hg), and of dLVP/dt at R45 min (1740.2+/-111.5 mm Hg.s-1 vs. 548.7+/-82.2 mm Hg.s-1) and R60 min (1199.8+/-104.6 mm Hg.s-1 vs. 466.9+/-61.1 mm Hg.s-1). The transgenic group also displayed development of less oxidative stress, decreased extent of infarction, and attenuated cardiomyocyte apoptotic cell death. Transgene overexpression of alphaB crystallin was therefore successful in diminishing the independent contributory effects of both necrosis and apoptosis on I/R-induced cell death. PMID:11156955

Ray, P S; Martin, J L; Swanson, E A; Otani, H; Dillmann, W H; Das, D K

2001-02-01

319

Potential transgenic routes to increase tree biomass.  

UK PubMed Central (United Kingdom)

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.

Dubouzet JG; Strabala TJ; Wagner A

2013-11-01

320

Possibility of target gene introgression from transgenic wheat into non-transgenic plants through pollens  

UK PubMed Central (United Kingdom)

The pollens of wheat could spread and set seeds in any direction at distances within 10m, and no seed set at distances out of 80m when castrated wheat was used as pollen recipient. The highest intraspecific rate of cross-pollination was only 0.24% within 1m, and spontaneous hybrids between Aegilops cylindrica L or Aegilops ovata L. and T aestivum L could be obtained at the rate of 0.25% and 0 respectively if non-castrated wheat, A cylindrica L and A ovata L were used as pollen recipients. For grain production, the frequency of gene introgression was much lower than that allowed by international standards even if the transgenic and non-transgenic wheat were planted nearby. When releasing transgenic wheat, those species that hybridize easily with common wheat could be considered as risky targets in its origin centers or its major distribution areas.

Lu Aizhi; Zhao He; Wang Tianyu; Wang Haibo

2002-01-01

 
 
 
 
321

In situ methods to localize transgenes and transcripts in interphase nuclei: a tool for transgenic plant research  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Genetic engineering of commercially important crops has become routine in many laboratories. However, the inability to predict where a transgene will integrate and to efficiently select plants with stable levels of transgenic expression remains a limitation of this technology. Fluorescence in situ hybridization (FISH) is a powerful technique that can be used to visualize transgene integration sites and provide a better understanding of transgene behavior. Studies using FISH to characterize transgene integration have focused primarily on metaphase chromosomes, because the number and position of integration sites on the chromosomes are more easily determined at this stage. However gene (and transgene) expression occurs mainly during interphase. In order to accurately predict the activity of a transgene, it is critical to understand its location and dynamics in the three-dimensional interphase nucleus. We and others have developed in situ methods to visualize transgenes (including single copy genes) and their transcripts during interphase from different tissues and plant species. These techniques reduce the time necessary for characterization of transgene integration by eliminating the need for time-consuming segregation analysis, and extend characterization to the interphase nucleus, thus increasing the likelihood of accurate prediction of transgene activity. Furthermore, this approach is useful for studying nuclear organization and the dynamics of genes and chromatin.

Santos Ana; Wegel Eva; Allen George C; Thompson William F; Stoger Eva; Shaw Peter; Abranches Rita

2006-01-01

322

Comparing the Agronomic and Grain Quality Characteristics of Transgenic Rice Lines Expressing cry1Ab vs. Non-Transgenic Controls  

Directory of Open Access Journals (Sweden)

Full Text Available This study aimed to investigate and compare the agronomic and grain quality attributes of three advanced backcross-derived transgenic rice lines expressing synthetic cry1Ab gene vs. non-transgenic control in a Randomized Complete Block Design (RCBD) under field conditions. The data exhibited that transgenic rice lines, Neda and Nemat were higher in height, earlier in maturity and highly resistant to striped stem borer (Chilo suppressalis) in comparing with non-transgenic varieties. In contrast, no significant difference was observed for transgenic Khazar as compared to its control, except for 1000-grain weight. Laboratory tests for grain physicochemical properties showed no significant variations between transgenic lines and non-transgenic controls. However, some variations for traits like Amylose Content (AC) and Gel Consistency (GC) were seen for transgenic Neda and Khazar, respectively. As regards the rice striped stem borer natural infestation, field-release experiment indicated that all three transgenic rice lines conferred a very high degree of resistance to rice striped stem borer as compared to non-transgenic check varieties.

G. Kiani; G.A. Nematzadeh; B. Ghareyazie; M. Sattari

2009-01-01

323

[Ecological risk of Bt transgenic cotton and its management strategy].  

UK PubMed Central (United Kingdom)

There may be several ecological impacts induced by transgenic cottons, apart from their direct impact on target pest. The interactions between target insect and transgenic cotton, and the way of toxic expressed by transgenic cotton varied with plant spatial parts and different growing stages are regarded as the main cause for insect to develop resistance. In transgenic cotton field, although chemicals applied to control major insect pest could be reduced greatly, insect community structure including insect pests and beneficial organisms is less stable than that of regular cotton field. It is much easier for minor insect pests developing to be major pest. In order to full utilization of transgenic cottons and keep their efficiency on target pest, methods including integrated pest management and breeding higher efficiency transgenic cottons by genetic engineering are proposed for management of pest resistance and non-target pests.

Ma J; Gao B; Wan F; Guo J

2003-03-01

324

[Ecological risk of Bt transgenic cotton and its management strategy].  

Science.gov (United States)

There may be several ecological impacts induced by transgenic cottons, apart from their direct impact on target pest. The interactions between target insect and transgenic cotton, and the way of toxic expressed by transgenic cotton varied with plant spatial parts and different growing stages are regarded as the main cause for insect to develop resistance. In transgenic cotton field, although chemicals applied to control major insect pest could be reduced greatly, insect community structure including insect pests and beneficial organisms is less stable than that of regular cotton field. It is much easier for minor insect pests developing to be major pest. In order to full utilization of transgenic cottons and keep their efficiency on target pest, methods including integrated pest management and breeding higher efficiency transgenic cottons by genetic engineering are proposed for management of pest resistance and non-target pests. PMID:12836558

Ma, Jun; Gao, Bida; Wan, Fanghao; Guo, Jianying

2003-03-01

325

Transgene introgression in crop relatives: molecular evidence and mitigation strategies.  

UK PubMed Central (United Kingdom)

Incorporation of crop genes into wild and weedy relative populations (i.e. introgression) has long been of interest to ecologists and weed scientists. Potential negative outcomes that result from crop transgene introgression (e.g. extinction of native wild relative populations; invasive spread by wild or weedy hosts) have not been documented, and few examples of transgene introgression exist. However, molecular evidence of introgression from non-transgenic crops to their relatives continues to emerge, even for crops deemed low-risk candidates for transgene introgression. We posit that transgene introgression monitoring and mitigation strategies are warranted in cases in which transgenes are predicted to confer selective advantages and disadvantages to recipient hosts. The utility and consequences of such strategies are examined, and future directions provided.

Kwit C; Moon HS; Warwick SI; Stewart CN Jr

2011-06-01

326

An efficient grafting technique for recovery of transgenic cotton plants.  

UK PubMed Central (United Kingdom)

Recovery of transgenic cotton plants from tissue culture condition to greenhouse condition is a critical step for improving cotton through genetic engineering. Traditional methods always cause low survival rate of transplanted plants. In 1998, we developed an efficient grafting technique for recovery of transgenic cotton plants, which significantly increased the survival rate of the transplanting regeneration plants. In this chapter, we present a detailed protocol for grafting transgenic cotton plants obtaining somatic embryogenesis.

Wang M; Wang Q; Zhang B

2013-01-01

327

Transgenic crops and sustainable development of agriculture in China  

UK PubMed Central (United Kingdom)

The transgenic crops are the biggest production of biotechnology on the agriculture.This paper starts from several problems during the new phase of agriculture development in China.Through the analysis of the value in use and security of transgenic crops,the authopr thinks the planting and developmnet of the transgeniccrops make for the sustainable development of agriculture in China.At addition,the prospect of transgenic crops and foods has been estimated.

2004-01-01

328

Expression of chicken liver cell adhesion molecule fusion genes in transgenic mice.  

Science.gov (United States)

The tissue-specific expression of the chicken liver cell adhesion molecule (L-CAM) was studied by generating transgenic mice. The rat insulin II promoter was fused to a chicken L-CAM cDNA or to chicken genomic L-CAM sequences. Mice carrying the cDNA showed no expression of L-CAM. Mice carrying L-CAM genomic sequences showed expression in the beta cells of the pancreas, suggesting that sequences in introns or in flanking regions are required for expression. Murine L-CAM was undetectable in the beta cells of the pancreas of those transgenic mice expressing chicken L-CAM and thus appeared to be down-regulated, but expression of the mouse protein was not altered at other sites. Chicken L-CAM was also found in extrapancreatic tissues such as skin, kidney, liver, lung, intestine, blood vessels, and the choroid plexus and leptomeninges of the central nervous system. These findings raised the possibility that the chicken L-CAM gene contains cis regulatory elements that interfere with the specificity of a tissue-specific promoter such as the rat insulin promoter. To test this hypothesis, transgenic mice were produced with a construct containing the murine neurofilament promoter fused to genomic chicken L-CAM sequences. Chicken L-CAM was expressed in the brain and spinal cord, where L-CAM is not normally found, but it was also found in some nonneural tissues (kidney, liver, intestine, lung) in which L-CAM is normally expressed. The combined results suggest that tissue-specific cis-acting elements in the chicken L-CAM gene, when combined with heterologous promoters/enhancers, can generate novel patterns of gene expression. Images

Begemann, M; Tan, S S; Cunningham, B A; Edelman, G M

1990-01-01

329

Conditional and targeted overexpression of vascular chymase causes hypertension in transgenic mice.  

Science.gov (United States)

We cloned a rat vascular chymase (RVCH) from smooth muscle cells (SMCs) that converts angiotensin I to II and is up-regulated in SMC from spontaneously hypertensive vs. normotensive rats. To determine whether increased activity of RVCH is sufficient to cause hypertension, transgenic mice were generated with targeted conditional expression of RVCH to SMC, with the use of the tetracycline-controlled transactivator (tTA). We confirmed conditional expression of RVCH by mRNA, protein, and chymase activity in the absence, but not in the presence, of dietary doxycycline. The systolic blood pressure (mmHg), measured by carotid artery cannulation at 10-12 weeks of age, was higher in tTA+/RVCH+ mice than in nonbinary transgenic littermates (136 +/- 4 vs. 109 +/- 3) (P metacholine-induced vasodilatation when compared with littermate controls or with the doxycyline-treated group. Our studies suggest that up-regulation of this vascular chymase is sufficient to cause a hypertensive arteriopathy, and that RVCH may be a candidate gene and a therapeutic target in patients with high blood pressure. PMID:11416217

Ju, H; Gros, R; You, X; Tsang, S; Husain, M; Rabinovitch, M

2001-06-19

330

Hepatic steatosis in transgenic mice overexpressing human histone deacetylase 1  

International Nuclear Information System (INIS)

[en] 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

331

Genetic background modifies neurodegeneration and neuroinflammation driven by misfolded human tau protein in rat model of tauopathy: implication for immunomodulatory approach to Alzheimer's disease  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Numerous epidemiological studies demonstrate that genetic background modifies the onset and the progression of Alzheimer's disease and related neurodegenerative disorders. The efficacious influence of genetic background on the disease pathway of amyloid beta has been meticulously described in rodent models. Since the impact of genetic modifiers on the neurodegenerative and neuroinflammatory cascade induced by misfolded tau protein is yet to be elucidated, we have addressed the issue by using transgenic lines expressing the same human truncated tau protein in either spontaneously hypertensive rat (SHR) or Wistar-Kyoto (WKY) genetic background. Methods Brains of WKY and SHR transgenic rats in the terminal stage of phenotype and their age-matched non-transgenic littermates were examined by means of immunohistochemistry and unbiased stereology. Basic measures of tau-induced neurodegeneration (load of neurofibrillary tangles) and neuroinflammation (number of Iba1-positive microglia, their activated morphology, and numbers of microglia immunoreactive for MHCII and astrocytes immunoreactive for GFAP) were quantified with an optical fractionator in brain areas affected by neurofibrillary pathology (pons, medulla oblongata). The stereological data were evaluated using two-way ANOVA and Student's t-test. Results Tau neurodegeneration (neurofibrillary tangles (NFTs), axonopathy) and neuroinflammation (microgliosis, astrocytosis) appeared in both WKY and SHR transgenic rats. Although identical levels of transgene expression in both lines were present, terminally-staged WKY transgenic rats displayed significantly lower final NFT loads than their SHR transgenic counterparts. Interestingly, microglial responses showed a striking difference between transgenic lines. Only 1.6% of microglia in SHR transgenic rats expressed MHCII in spite of having a robust phagocytic phenotype, whereas in WKY transgenic rats, 23.2% of microglia expressed MHCII despite displaying a considerably lower extent of transformation into phagocytic phenotype. Conclusions These results show that the immune response represents a pivotal and genetically variable modifying factor that is able to influence vulnerability to neurodegeneration. Therefore, targeted immunomodulation could represent a prospective therapeutic approach to Alzheimer's disease.

Stozicka Zuzana; Zilka Norbert; Novak Petr; Kovacech Branislav; Bugos Ondrej; Novak Michal

2010-01-01

332

An industry perspective on the utility of short-term carcinogenicity testing in transgenic mice in pharmaceutical development.  

Science.gov (United States)

International guidelines allow for use of a short-term cancer bioassay (twenty-six weeks) in transgenic mice as a substitute for one of the two required long-term rodent bioassays in the preclinical safety evaluation of pharmaceuticals. The two models that have gained the widest acceptance by sponsors and regulatory authorities are the CB6F1-RasH2 mouse hemizygous for a human H-ras transgene and the B6.129N5-Trp53 mouse heterozygous for a p53 null allele. The p53(+/-) model is of particular value for compounds with residual concern that genotoxic activity may contribute to tumorigenesis. The rasH2 model is an appropriate alternative without regard to evidence of genotoxic potential. Since results from a short-term bioassay can be obtained relatively early in drug development, there is the potential for more timely assessment of cancer risk for individuals in long-term clinical trials. Use of these models in preclinical safety evaluation also significantly reduces animal use, time, and manpower. Preliminary findings indicate that prediction of two-year rat bioassay outcomes based on data from chronic rat toxicity studies, together with early assessment of carcinogenic potential in short-term transgenic models, may have the potential to increase the timeliness and efficiency of strategies for the identification of human carcinogenic hazards. PMID:19893055

Storer, Richard D; Sistare, Frank D; Reddy, M Vijayaraj; DeGeorge, Joseph J

2009-11-05

333

An industry perspective on the utility of short-term carcinogenicity testing in transgenic mice in pharmaceutical development.  

UK PubMed Central (United Kingdom)

International guidelines allow for use of a short-term cancer bioassay (twenty-six weeks) in transgenic mice as a substitute for one of the two required long-term rodent bioassays in the preclinical safety evaluation of pharmaceuticals. The two models that have gained the widest acceptance by sponsors and regulatory authorities are the CB6F1-RasH2 mouse hemizygous for a human H-ras transgene and the B6.129N5-Trp53 mouse heterozygous for a p53 null allele. The p53(+/-) model is of particular value for compounds with residual concern that genotoxic activity may contribute to tumorigenesis. The rasH2 model is an appropriate alternative without regard to evidence of genotoxic potential. Since results from a short-term bioassay can be obtained relatively early in drug development, there is the potential for more timely assessment of cancer risk for individuals in long-term clinical trials. Use of these models in preclinical safety evaluation also significantly reduces animal use, time, and manpower. Preliminary findings indicate that prediction of two-year rat bioassay outcomes based on data from chronic rat toxicity studies, together with early assessment of carcinogenic potential in short-term transgenic models, may have the potential to increase the timeliness and efficiency of strategies for the identification of human carcinogenic hazards.

Storer RD; Sistare FD; Reddy MV; DeGeorge JJ

2010-01-01

334

The transgenic window on lymphoid malignancy.  

UK PubMed Central (United Kingdom)

Transgenic mice bearing an oncogene targetted for expression in a specific tissue can reveal how that oncogene influences differentiation and help to delineate the pathways to malignancy. To explore lymphoid neoplasia, we have made strains of transgenic mice bearing different oncogenes driven by the immunoglobulin heavy chain enhancer (E mu), which promotes expression within lymphocytes and certain myeloid cells. The prototype E mu-myc mice succumb to pre-B and B cell lymphomas, following a preneoplastic phase in which cycling pre-B cells are overproduced. The similar fate of E mu-N-myc mice suggests that N-myc and myc have overlapping functions. Surprisingly, E mu-N-ras mice develop T lymphomas and macrophage tumours but no B lineage tumours; thus the ability of ras to initiate tumorigenesis may be lineage specific. Similarly, the high predisposition of E mu-v-abl mice to develop plasmacytomas may indicate that v-abl is oncogenic only at certain stages of B cell maturation. The bcl-2 gene promotes cell survival rather than proliferation, and E mu-bcl-2 mice produce copious resting B lymphocytes. The random onset and monoclonality of tumours in the transgenic strains argues for spontaneous genetic alterations that cooperate with the trans-oncogene. Indeed, most plasmacytomas of E mu-v-abl mice bear spontaneous myc rearrangements. Moreover, a minority of E mu-myc B lymphomas exhibit ras mutation, and the tumorigenesis can be reconstructed by crossing E mu-myc and E mu-ras mice, or by retroviral delivery of v-ras or v-raf, either in vitro or in vivo. To access novel cooperating oncogenes, we are using a retrovirus lacking an oncogene as an insertional mutagen. This approach should be applicable to any trans-oncogenic strain and help to delineate the genetic events that trigger malignant clones.

Adams JM; Harris AW; Vaux DL; Alexander WS; Rosenbaum H; Klinken SP; Strasser A; Bath ML; McNeall J; Cory S

1989-01-01

335

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

UK PubMed Central (United Kingdom)

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.

Ripoll CB; Bunnell BA

2009-01-01

336

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.

Ripoll Cynthia B; Bunnell Bruce A

2009-01-01

337

Recent advances in the development of new transgenic animal technology.  

UK PubMed Central (United Kingdom)

Transgenic animal technology is one of the fastest growing biotechnology areas. It is used to integrate exogenous genes into the animal genome by genetic engineering technology so that these genes can be inherited and expressed by offspring. The transgenic efficiency and precise control of gene expression are the key limiting factors in the production of transgenic animals. A variety of transgenic technologies are available. Each has its own advantages and disadvantages and needs further study because of unresolved technical and safety issues. Further studies will allow transgenic technology to explore gene function, animal genetic improvement, bioreactors, animal disease models, and organ transplantation. This article reviews the recently developed animal transgenic technologies, including the germ line stem cell-mediated method to improve efficiency, gene targeting to improve accuracy, RNA interference-mediated gene silencing technology, zinc-finger nuclease gene targeting technology and induced pluripotent stem cell technology. These new transgenic techniques can provide a better platform to develop transgenic animals for breeding new animal varieties and promote the development of medical sciences, livestock production, and other fields.

Miao X

2013-03-01

338

Airway Specific Inducible Transgene Expression Using Aerosolized Doxycycline.  

UK PubMed Central (United Kingdom)

Tissue specific transgene expression using tet-regulated promoter/operator elements has been used to revolutionize our understanding of cellular and molecular processes. However, since most tet-regulated mouse strains use promoters that are expressed in multiple tissues, it is often impossible to achieve an organ-specific effect. Indeed, in the extreme case, unwanted transgene expression in other organ systems causes lethality that precludes the study of the transgene in the actual organ of interest. Here, we describe a novel approach to activate tet-inducible transgene expression solely in the airway by using aerosolized doxycycline administration. By optimizing the dosage and duration of aerosolized doxycycline exposure in mice possessing a ubiquitously expressed Rosa26 promoter-driven rtTA element, we induce transgene expression exclusively in the airways. We detect no changes in the cellular composition or proliferative behavior of the airway cells. We used this newly developed method to achieve airway basal stem cell-specific transgene expression using a KRT5-driven rtTA driver line to induce Notch pathway activation. We observed an expected, but more robust, mucous metaplasia phenotype than in animals given doxycycline systemically. Additionally, phenotypes outside of the lung that were evident when doxycycline was given systemically were now absent. Thus, our approach allows rapid and efficient airway-specific transgene expression. A suite of pre-existing transgenic mice can now be used to specifically study airway biology.

Tata PR; Pardo-Saganta A; Prabhu M; Vinarsky V; Law BM; Fontaine BA; Tager AM; Rajagopal J

2013-07-01

339

Transgenic Crops and Sustainable Agriculture in the European Context  

Science.gov (United States)

The rapid adoption of transgenic crops in the United States, Argentina, and Canada stands in strong contrast to the situation in the European Union (EU), where a de facto moratorium has been in place since 1998. This article reviews recent scientific literature relevant to the problematic introduction of transgenic crops in the EU to assess if…

Ponti, Luigi

2005-01-01

340

Resistance of transgenic watermelon (Citrullus vulgaris Schrad) to virus disease  

UK PubMed Central (United Kingdom)

This paper reports the results of the resistance of transgenic watermelon (Da Zheng Bao) to virus disease. The results show that whether in green-room or in field, compared with the control groups, transgenic plants can delay the disease infection and reduce the incidence and the symptoms of virus disease, thus showing a certain resistance to relevant viruses.

Wang Huizhong; Zhao Peijie; Zhou Xiaoyun

1998-12-01

 
 
 
 
341

Zebrafish transgenic Enhancer TRAP line database (ZETRAP)  

Directory of Open Access Journals (Sweden)

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.

Choo Benjamin GH; Kondrichin Igor; Parinov Sergey; Emelyanov Alexander; Go William; Toh Wei-chang; Korzh Vladimir

2006-01-01

342

Human monoclonal antibodies from transgenic mice.  

UK PubMed Central (United Kingdom)

Since the 1986 regulatory approval of muromonomab-CD3, a mouse monoclonal antibody (MAb) directed against the T cell CD3epsilon antigen, MAbs have become an increasingly important class of therapeutic compounds in a variety of disease areas ranging from cancer and autoimmune indications to infectious and cardiac diseases. However, the pathway to the present acceptance of therapeutic MAbs within the pharmaceutical industry has not been smooth. A major hurdle for antibody therapeutics has been the inherent immunogenicity of the most readily available MAbs, those derived from rodents. A variety of technologies have been successfully employed to engineer MAbs with reduced immunogenicity. Implementation of these antibody engineering technologies involves in vitro optimization of lead molecules to generate a clinical candidate. An alternative technology, involving the engineering of strains of mice to produce human instead of mouse antibodies, has been emerging and evolving for the past two decades. Now, with the 2006 US regulatory approval of panitumumab, a fully human antibody directed against the epidermal growth factor receptor, transgenic mice expressing human antibody repertoires join chimerization, CDR grafting, and phage display technologies, as a commercially validated antibody drug discovery platform. With dozens of additional transgenic mouse-derived human MAbs now in clinical development, this new drug discovery platform appears to be firmly established within the pharmaceutical industry.

Lonberg N

2008-01-01

343

Investigations into the hypothesis of transgenic cannabis.  

UK PubMed Central (United Kingdom)

The unusual concentration of cannabinoids recently found in marijuana samples submitted to the forensic laboratory for chemical analysis prompted an investigation into whether genetic modifications have been made to the DNA of Cannabis sativa L. to increase its potency. Traditional methods for the detection of genetically modified organisms (GMO) were used to analyze herbal cannabis preparations. Our analyses support the hypothesis that marijuana samples submitted to forensic laboratories and characterized by an abnormal level of ?(9)-THC are the product of breeding selection rather than of transgenic modifications. Further, this research has shown a risk of false positive results associated with the poor quality of the seized samples and probably due to the contamination by other transgenic vegetable products. On the other hand, based on these data, a conclusive distinction between the hypothesis of GMO plant contamination and the other of genetic modification of cannabis cannot be made requiring further studies on comparative chemical and genetic analyses to find out an explanation for the recently detected increased potency of cannabis.

Cascini F

2012-05-01

344

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.

1995-01-01

345

[Biofuels, food security and transgenic crops].  

UK PubMed Central (United Kingdom)

Soaring global food prices are threatening to push more poor people back below the poverty line; this will probably become aggravated by the serious challenge that increasing population and climate changes are posing for food security. There is growing evidence that human activities involving fossil fuel consumption and land use are contributing to greenhouse gas emissions and consequently changing the climate worldwide. The finite nature of fossil fuel reserves is causing concern about energy security and there is a growing interest in the use of renewable energy sources such as biofuels. There is growing concern regarding the fact that biofuels are currently produced from food crops, thereby leading to an undesirable competition for their use as food and feed. Nevertheless, biofuels can be produced from other feedstocks such as lingo-cellulose from perennial grasses, forestry and vegetable waste. Biofuel energy content should not be exceeded by that of the fossil fuel invested in its production to ensure that it is energetically sustainable; however, biofuels must also be economically competitive and environmentally acceptable. Climate change and biofuels are challenging FAO efforts aimed at eradicating hunger worldwide by the next decade. Given that current crops used in biofuel production have not been domesticated for this purpose, transgenic technology can offer an enormous contribution towards improving biofuel crops' environmental and economic performance. The present paper critically presents some relevant relationships between biofuels, food security and transgenic plant technology.

Acosta O; Chaparro-Giraldo A

2009-03-01

346

Process for obtaining antimycosis cut flower flameray gerbera by transgene  

UK PubMed Central (United Kingdom)

The invention relates to a method for transgenically obtaining a nosomycosis resistant cut gerbera flower. The method obtains a transgenic plant through the following steps of: separating an antifungal gene, namely a chitinase gene Chi and a beta-1, 3-dextranase gene Glu, cloning the Chi and the Glu, constructing a bivalent expression vector, constructing a high-efficient genetic transformation regeneration system of the cut gerbera flower, transformation of the cut gerbera flower by the antifungal gene and so on. The method uses CaMV35S as starter and transforms genes like Chi, Glu, Npt II and so on into a cut gerbera flower variety 'redcap' and then obtains the transgenic plant, thereby providing technical support for obtaining a large number of antifungal transgenic African daisy strains. The method effectively provides the antifungal transgenic African daisy strains and simultaneously provides a quick, effective and stable technical path for African daisy genetic engineering breeding.

HAN LI; HUIJUN YAN; TING ZHANG; SHENCHONG LI; JIHUA WANG; HAO ZHANG; XIA LI

347

Advancing environmental risk assessment for transgenic biofeedstock crops  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Transgenic modification of plants is a key enabling technology for developing sustainable biofeedstocks for biofuels production. Regulatory decisions and the wider acceptance and development of transgenic biofeedstock crops are considered from the context of science-based risk assessment. The risk assessment paradigm for transgenic biofeedstock crops is fundamentally no different from that of current generation transgenic crops, except that the focus of the assessment must consider the unique attributes of a given biofeedstock crop and its environmental release. For currently envisioned biofeedstock crops, particular emphasis in risk assessment will be given to characterization of altered metabolic profiles and their implications relative to non-target environmental effects and food safety; weediness and invasiveness when plants are modified for abiotic stress tolerance or are domesticated; and aggregate risk when plants are platforms for multi-product production. Robust risk assessments for transgenic biofeedstock crops are case-specific, initiated through problem formulation, and use tiered approaches for risk characterization.

Wolt Jeffrey D

2009-01-01

348

Treatment of Transgenic Crops with Mixtures of Fiproles and Chloronicotinyls  

UK PubMed Central (United Kingdom)

The Invention relates to methods for increasing the production potential of plants and/or controlling pests in plants with at least one transgenic modification related to yield increase as compared to a corresponding wild-type plant, comprising treating the location where the plant with at least one transgenic modification is growing or is expected to grow and/or the transgenic plant with at least one transgenic modification or propagation material of the plant with at least one transgenic modification with an effective amount of an insecticidal composition comprising a component A, selected from the group consisting of imidacloprid, thiacloprid, clothianidin, acetamiprid, dinotefuran, nitenpyram, and thiamethoxam and a component B, selected from the group consisting of fipronil and ethiprole.

ANDERSCH WOLFRAM; HUNGENBERG HEIKE; SPRINGER BERND; SACHAU STEFAN; ROOIJEN CASPER ISAAK

349

Improved neuronal transgene expression from an AAV-2 vector with a hybrid CMV enhancer/PDGF-beta promoter.  

UK PubMed Central (United Kingdom)

BACKGROUND: Adeno-associated virus type 2 (AAV-2) vectors are highly promising tools for gene therapy of neurological disorders. After accommodating a cellular promoter, AAV-2 vectors are able to drive sustained expression of transgene in the brain. This study aimed to develop AAV-2 vectors that also facilitate a high level of neuronal expression by enhancing the strength of a neuron-specific promoter, the human platelet-derived growth factor beta-chain (PDGF) promoter. METHODS AND RESULTS: A hybrid promoter approach was adopted to fuse the enhancer of human cytomegalovirus immediately early (CMV) promoter to the PDGF promoter. In cultured cortex neurons, AAV-2 vectors containing the hybrid promoter augmented transgene expression up to 20-fold over that mediated by titer-matched AAV-2 vectors with the PDGF promoter alone and 4-fold over the CMV enhancer/promoter. Injection of AAV-2 vectors with the hybrid promoter into the rat striatum resulted in neuron-specific transgene expression, the level of which was about 10-fold higher than those provided by the two control AAV-2 expression cassettes at 4 weeks post-injection and maintained for at least 12 weeks. Gene expression in the substantia nigra through possible retrograde transport of the AAV-2 vectors injected into the striatum was not obvious. After direct injection of AAV-2 vectors into the substantia nigra, transgene expression driven by the hybrid promoter was observed specifically in dopaminergic neurons and its level was about 3 and 17 times higher than that provided by the PDGF promoter alone and the CMV enhancer/promoter, respectively. CONCLUSIONS: Enhanced transgene capacity plus neuron-specificity of the AAV-2 vectors developed in this study might prove valuable for gene therapy of Parkinson's disease.

Wang CY; Guo HY; Lim TM; Ng YK; Neo HP; Hwang PY; Yee WC; Wang S

2005-07-01

350

Functional conservation between rodents and chicken of regulatory sequences driving skeletal muscle gene expression in transgenic chickens  

Directory of Open Access Journals (Sweden)

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.

McGrew Michael J; Sherman Adrian; Lillico Simon G; Taylor Lorna; Sang Helen

2010-01-01

351

Illegal gene flow from transgenic creeping bentgrass: the saga continues.  

UK PubMed Central (United Kingdom)

Ecologists have paid close attention to environmental effects that fitness-enhancing transgenes might have following crop-to-wild gene flow (e.g. Snow et al. 2003). For some crops, gene flow also can lead to legal problems,especially when government agencies have not approved transgenic events for unrestricted environmental release.Creeping bentgrass (Agrostis stolonifera), a common turf grass used in golf courses, is the focus of both areas of concern. In 2002, prior to expected deregulation (still pending), The Scotts Company planted creeping bentgrass with transgenic resistance to the herbicide glyphosate,also known as RoundUp, on 162 ha in a designated control area in central Oregon (Fig. 1).Despite efforts to restrict gene flow, wind-dispersed pollen carried transgenes to florets of local A. stolonifera and A. gigantea as far as 14 km away, and to sentinel plants placed as far as 21 km away (Watrud et al. 2004).Then, in August 2003, a strong wind event moved transgenic seeds from wind rows of cut bentgrass into nearby areas. The company’s efforts to kill all transgenic survivors in the area failed: feral glyphosate-resistant populations of A. stolonifera were found by Reichman et al.(2006), and 62% of 585 bentgrass plants had the telltale CP4 EPSPS transgene in 2006 (Zapiola et al. 2008; Fig. 2).Now, in this issue, the story gets even more interesting as Zapiola & Mallory-Smith (2012) describe a transgenic,intergeneric hybrid produced on a feral, transgenic creeping bentgrass plant that received pollen from Polypogon monspeliensis (rabbitfoot grass). Their finding raises a host of new questions about the prevalence and fitness of intergeneric hybrids, as well as how to evaluate the full extent of gene flow from transgenic crops.

Snow AA

2012-10-01

352

The use of yeast artificial chromosomes in transgenic animals: expression studies of the tyrosinase gene in transgenic mice.  

Science.gov (United States)

Variegation and inherited somatic mosaicism has been observed in transgenic mice carrying yeast artificial chromosomes (YACs) in which a DNAse I hypersensitive site (HS) located -12 kb upstream of the mouse tyrosinase gene had been deleted. At present, we are generating new transgenic animals with minor deletions of the HS. PMID:10596759

Giraldo, P; Giménez, E; Montoliu, L

1999-11-01

353

Comparing the Agronomic and Grain Quality Characteristics of Transgenic Rice Lines Expressing cry1Ab vs. Non-Transgenic Controls  

Digital Repository Infrastructure Vision for European Research (DRIVER)

This study aimed to investigate and compare the agronomic and grain quality attributes of three advanced backcross-derived transgenic rice lines expressing synthetic cry1Ab gene vs. non-transgenic control in a Randomized Complete Block Design (RCBD) under field conditions. The data exhibi...

G. Kiani; G.A. Nematzadeh; B. Ghareyazie; M. Sattari

354

High-value products from transgenic maize.  

UK PubMed Central (United Kingdom)

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.

Naqvi S; Ramessar K; Farré G; Sabalza M; Miralpeix B; Twyman RM; Capell T; Zhu C; Christou P

2011-01-01

355

Magnetic biomineralisation in Huntington's disease transgenic mice  

International Nuclear Information System (INIS)

[en] 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

356

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