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1.
J Back Musculoskelet Rehabil ; 31(2): 221-229, 2018.
Article in English | MEDLINE | ID: mdl-29154263

ABSTRACT

BACKGROUND: Ankle sprains are common musculoskeletal injuries in which the ligaments of the ankle partially or completely tear due to sudden stretching. OBJECTIVES: To critically appraise, evaluate and establish the best available evidence to determine the effectiveness of proprioceptive and neuromuscular training (PNT) compared to bracing in reducing the recurrence rate of ankle sprains in athletes. METHODOLOGY: The following seven databases were searched in June 2017: PubMed, Cochrane Library, PEDro, ScienceDirect, Scopus, SPORTDiscus, EBSCO Host: CINAHL. The main search terms used were "ankle sprains", "proprioceptive training", "neuromuscular training" and "bracing". The quality of the trials were critically appraised according to the PEDro scale. The RevMan 5© software was used to pool results. RESULTS: Three studies met the inclusion criteria and the quality according to the PEDro scale ranged from 4/10-7/10. The pooled data showed no difference between PNT and bracing in reducing the recurrence rate of ankle sprains in athletes at 12 months after initiation of the study. CONCLUSION: This systematic review of the overall effect suggested that current evidence (Level II) does not favour the use of PNT over bracing in reducing the recurrence rate of ankle sprains. Physiotherapists are advised to use either PNT or bracing according to the patients preference and their own expertise.


Subject(s)
Ankle Injuries/prevention & control , Athletic Injuries/prevention & control , Braces , Physical Therapy Modalities , Proprioception , Sprains and Strains/prevention & control , Humans , Recurrence
2.
JPEN J Parenter Enteral Nutr ; 25(4): 194-202, 2001.
Article in English | MEDLINE | ID: mdl-11434650

ABSTRACT

BACKGROUND: Hypertension in kidney transplant (KT) patients may result from attenuated whole-body nitric oxide (NO) content and abnormal NO-mediated vasodilation. Increasing NO bioavailability with L-arginine (ARG) could theoretically restore the NO-mediated vasodilatory response and lower blood pressure. METHODS: In a prospective pilot study, 6 normotensive volunteers and 10 KT patients received oral supplements of ARG (9.0 g/d) for 9 days, then 18.0 g/d for 9 more days. Six hemodialysis (HD) and 4 peritoneal dialysis patients received the same dose for 14 days. Five KT patients received 30 mL/d of canola oil (CanO) in addition to ARG. Systolic (SBP) and diastolic (DBP) blood pressure, creatinine clearance (CCr), and serum creatinine (Cr) were measured at baseline, day 9, and day 18. In a subsequent study, 20 hypertensive KT patients with stable but abnormal renal function were randomized in a crossover study to start ARG-only or ARG+CanO supplements for two 2-month periods with an intervening month of no supplementation. SBP, DBP, CCr, and Cr were measured monthly for 7 months. RESULTS: In the pilot study, ARG reduced the SBP in HD patients from 171.5 +/- 7.5 mmHg (baseline) to 142.8 +/- 8.3 mmHg (p = .028). In the crossover study, SBP was reduced from baseline (155.9 +/- 5.0 mmHg), after the first 2 months (143.2 +/- 3.2 mmHg; p = .03) and subsequent 2 months (143.3 +/- 2.5 mmHg; p = .014) of supplementation. DBP was also reduced after supplementation in both studies. CanO had no effect on blood pressure. Renal function did not change. CONCLUSIONS: Oral preparations of ARG (+/-CanO) were well tolerated for up to 60 consecutive days and had favorable effects on SBP and DBP in hypertensive KT and HD patients.


Subject(s)
Arginine/administration & dosage , Blood Pressure/drug effects , Hypertension/drug therapy , Kidney Diseases/therapy , Kidney Transplantation , Renal Dialysis , Adult , Arginine/therapeutic use , Blood Pressure/physiology , Creatinine/blood , Creatinine/urine , Cross-Over Studies , Dietary Supplements , Fatty Acids, Monounsaturated/administration & dosage , Female , Humans , Kidney Diseases/complications , Longitudinal Studies , Male , Middle Aged , Nitrates/blood , Nitric Oxide/metabolism , Pilot Projects , Prospective Studies , Rapeseed Oil , Vasodilation
4.
Hum Gene Ther ; 11(3): 415-27, 2000 Feb 10.
Article in English | MEDLINE | ID: mdl-10697116

ABSTRACT

Liver toxicity and inflammation were assessed in C57BL/6, CBA, and BALB/c mice injected intravenously with a series of recombinant adenoviruses deleted simultaneously in E1/E3, in E1/E3/E2A, or in E1/E3/E4. All vectors were either devoid of transgenes or carried in E1 the human CFTR cDNA under the control of the CMV promoter. Injection of the E1/E3-deleted vector induced a significant liver dystrophy and inflammatory responses that were accompanied by an increased serum transaminase concentration. The vector toxicity remained elevated on additional deletion of the E2A gene and was further enhanced when hCFTR was expressed. In contrast, additional deletion of E4 led to a reduction in hepatotoxicity, suggesting an active role of E4 gene products in liver injury. However, deletion of E4 also led to a loss of transgene expression. To identify the individual E4 product(s) involved in liver toxicity and in the regulation of transgene expression, a series of isogenic E1/E3-deleted vectors, with or without the hCFTR transgene, and containing various combinations of functional E4 open reading frames (ORFs), were evaluated in vitro and in vivo. We demonstrate that liver injury was markedly reduced with vectors containing either ORF3 alone or ORF3,4 while vectors containing ORF4, ORF6,7 or ORF3,6,7 still displayed elevated hepatotoxicity and inflammatory responses. Moreover, transgene expression was restored when ORF3,4 or ORF3,6,7 was retained in the vector. These results highlight the importance of the E4 gene products in the design of improved in vivo gene transfer vectors.


Subject(s)
Adenoviridae/genetics , Adenovirus E4 Proteins/genetics , Chemical and Drug Induced Liver Injury/pathology , Gene Transfer Techniques , Liver/pathology , Animals , Chemical and Drug Induced Liver Injury/etiology , Cystic Fibrosis Transmembrane Conductance Regulator/genetics , Cystic Fibrosis Transmembrane Conductance Regulator/metabolism , Female , Gene Deletion , Gene Transfer Techniques/adverse effects , Genetic Vectors , Humans , Injections, Intravenous , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Inbred CBA , Open Reading Frames , Transgenes
5.
J Gene Med ; 2(6): 433-43, 2000.
Article in English | MEDLINE | ID: mdl-11199264

ABSTRACT

BACKGROUND: Strong and stable transgene expression is fundamental to the success of recombinant adenovirus vectors in human gene therapy. However, control of transgene expression is a complex process, involving both viral and cellular factors. In this study, the influence of the E4 adenoviral region on the activity of various promoters was investigated in vitro and in vivo. METHODS: Pairs of isogenic E1o and E1oE4o vectors were generated and compared. Levels of transgene expression were determined by Northern blot, ELISA and FACS analysis. Initiation of transcription was studied by nuclear run-on assays. RESULTS: Similar to the viral CMV and RSV promoters, the activity of the ubiquitous cellular PGK promoter required the presence of the E4 genes in vitro and in vivo. In contrast, transgene expression from selected liver- and tumor-specific promoters did not require E4 functions. CONCLUSION: Together with the reported low liver toxicity of E1oE4o vectors, the independence of E4 of liver-specific promoters renders such vectors interesting alternatives to the use of gutless vectors.


Subject(s)
Adenovirus E4 Proteins/genetics , Avian Sarcoma Viruses/genetics , Cytomegalovirus/genetics , Promoter Regions, Genetic , 3T3 Cells , Adenovirus E1 Proteins/genetics , Animals , COS Cells , Cell Line , Chlorocebus aethiops , Gene Deletion , Gene Expression Regulation , Gene Transfer Techniques , Genetic Vectors/genetics , HeLa Cells , Humans , Mice , Mice, Inbred Strains , Mice, SCID , Transcription, Genetic , Transgenes/genetics , Tumor Cells, Cultured , Vero Cells
6.
J Virol ; 73(10): 8308-19, 1999 Oct.
Article in English | MEDLINE | ID: mdl-10482581

ABSTRACT

In a previous study we showed that multiple deletions of the adenoviral regulatory E1/E3/E4 or E1/E3/E2A genes did not influence the in vivo persistence of the viral genome or affect the antiviral host immune response (Lusky et al., J. Virol. 72:2022-2032, 1998). In this study, the influence of the adenoviral E4 region on the strength and persistence of transgene expression was evaluated by using as a model system the human cystic fibrosis transmembrane conductance regulator (CFTR) cDNA transcribed from the cytomegalovirus (CMV) promoter. We show that the viral E4 region is indispensable for persistent expression from the CMV promoter in vitro and in vivo, with, however, a tissue-specific modulation of E4 function(s). In the liver, E4 open reading frame 3 (ORF3) was necessary and sufficient to establish and maintain CFTR expression. In addition, the E4 ORF3-dependent activation of transgene expression was enhanced in the presence of either E4 ORF4 or E4 ORF6 and ORF6/7. In the lung, establishment of transgene expression was independent of the E4 gene products but maintenance of stable transgene expression required E4 ORF3 together with either E4 ORF4 or E4 ORF6 and ORF6/7. Nuclear run-on experiments showed that initiation of transcription from the CMV promoter was severely reduced in the absence of E4 functions but could be partially restored in the presence of either ORF3 and ORF4 or ORFs 1 through 4. These results imply a direct involvement of some of the E4-encoded proteins in the transcriptional regulation of heterologous transgenes. We also report that C57BL/6 mice are immunologically weakly responsive to the human CFTR protein. This observation implies that such mice may constitute attractive hosts for the in vivo evaluation of vectors for cystic fibrosis gene therapy.


Subject(s)
Adenoviridae , Adenovirus E4 Proteins/genetics , Cystic Fibrosis Transmembrane Conductance Regulator/genetics , Gene Transfer Techniques , Genetic Vectors , Animals , Genetic Therapy , Humans , Mice , Mice, Inbred C57BL , Promoter Regions, Genetic , Transcription, Genetic
8.
Strahlenther Onkol ; 174 Suppl 3: 99-100, 1998 Nov.
Article in English | MEDLINE | ID: mdl-9830470

ABSTRACT

AIM: In many case reports the success of treatment of late complications of radiotherapy with hyperbaric oxygenation (HBO) has been shown. This synopsis attempts to review HBO in the treatment of chronic radiation injury of the bladder. PATIENTS AND METHODS: Three female patients who had developed urge-incontinence after a Wertheim operation and combined brachy-teletherapy with cobalt-60 and afterloading and did not respond to various drug therapies, were treated with HBO to a maximum of 40 applications. RESULTS: In all patients HBO altered and inverted the dynamic process underlying chronic bladder changes after irradiation. Rationales for the HBO are the reduction of tissue hypoxia and the induction of neoangiogenesis. CONCLUSIONS: There are no prospective trials up to date showing the benefit of HBO to urinary disorders caused by radiation cystitis. The positive results of our retrospective study should encourage clinicians to initiate prospective studies with the use of HBO in the treatment of radiation cystitis.


Subject(s)
Brachytherapy/adverse effects , Hyperbaric Oxygenation , Hysterectomy , Radiation Injuries/therapy , Radiotherapy/adverse effects , Urinary Bladder/radiation effects , Urinary Incontinence/therapy , Cobalt Radioisotopes/adverse effects , Combined Modality Therapy/adverse effects , Female , Humans , Radiation Injuries/etiology , Retrospective Studies , Urinary Bladder/physiopathology , Urinary Incontinence/etiology
9.
Strahlenther Onkol ; 174(9): 482-3, 1998 Sep.
Article in English | MEDLINE | ID: mdl-9765691

ABSTRACT

AIM: Our objective was to investigate the effectiveness of hyperbaric oxygenation (HBO) in the treatment of radiation proctitis. The current literature was reviewed with regard to the necessary number of HBO treatments. PATIENTS AND METHODS: Two patients with proctitis after pelvic irradiation were treated with 40 and 38 HBO treatments, respectively. Hyperbaric oxygenation was delivered at 240 kPa over 90 min. RESULTS: In one patient, proctosigmoidoscopy showed a significant improvement after 40 HBO sessions. The other patient interrupted therapy after 38 HBO treatments without subjective change. The reported number of HBO sessions for a successful treatment of radiation proctitis ranges from 12 to 90. CONCLUSION: HBO should be considered before more invasive treatment modalities are performed for radiation proctitis.


Subject(s)
Hyperbaric Oxygenation/methods , Proctitis/therapy , Radiotherapy/adverse effects , Aged , Female , Humans , Male , Proctitis/etiology
12.
J Virol ; 72(3): 2022-32, 1998 Mar.
Article in English | MEDLINE | ID: mdl-9499056

ABSTRACT

Isogenic, E3-deleted adenovirus vectors defective in E1, E1 and E2A, or E1 and E4 were generated in complementation cell lines expressing E1, E1 and E2A, or E1 and E4 and characterized in vitro and in vivo. In the absence of complementation, deletion of both E1 and E2A completely abolished expression of early and late viral genes, while deletion of E1 and E4 impaired expression of viral genes, although at a lower level than the E1/E2A deletion. The in vivo persistence of these three types of vectors was monitored in selected strains of mice with viral genomes devoid of transgenes to exclude any interference by immunogenic transgene-encoded products. Our studies showed no significant differences among the vectors in the short-term maintenance and long-term (4-month) persistence of viral DNA in liver and lung cells of immunocompetent and immunodeficient mice. Furthermore, all vectors induced similar antibody responses and comparable levels of adenovirus-specific cytotoxic T lymphocytes. These results suggest that in the absence of transgenes, the progressive deletion of the adenovirus genome does not extend the in vivo persistence of the transduced cells and does not reduce the antivirus immune response. In addition, our data confirm that, in the absence of transgene expression, mouse cellular immunity to viral antigens plays a minor role in the progressive elimination of the virus genome.


Subject(s)
Adenovirus E1 Proteins/genetics , Adenovirus E2 Proteins/genetics , Adenovirus E4 Proteins/genetics , Adenoviruses, Human , Capsid Proteins , Gene Deletion , Genetic Vectors , Adenovirus E1 Proteins/biosynthesis , Adenovirus E1 Proteins/immunology , Adenovirus E2 Proteins/biosynthesis , Adenovirus E2 Proteins/immunology , Adenovirus E4 Proteins/immunology , Adenoviruses, Human/genetics , Adenoviruses, Human/immunology , Adenoviruses, Human/metabolism , Animals , Capsid/biosynthesis , Cell Line , DNA-Binding Proteins/biosynthesis , Female , Genetic Vectors/genetics , Genetic Vectors/immunology , Genetic Vectors/metabolism , Genome, Viral , Humans , Mice , Mice, Inbred CBA , Mice, SCID , Time Factors , Virus Latency
13.
Immunol Lett ; 57(1-3): 19-25, 1997 Jun 01.
Article in English | MEDLINE | ID: mdl-9232420

ABSTRACT

E1, E3-deleted, replication-deficient recombinant adenoviruses are widely studied as vectors for their capacity to transfer therapeutic genes in vivo. They can infect a wide variety of dividing and quiescent cells from different organs and possess a large packaging capacity. One of the major limitations in the use of these vectors for gene therapy is the transient expression of the transgene in vivo and the poor transduction efficiency when re-administered. Despite the deletion of the viral E1 region, low level of early and late viral genes are expressed in vivo. Thus, viral antigens plus those derived from transgene expression in transduced cells contribute to cellular immune responses leading to the destruction of these cells. Production of anti-adenovirus antibodies, the cellular immune response as well as the early non-specific clearance of the vectors, constitute barriers to successful gene therapy. New vectors have been derived with additional deletions in the E2a or the E4 regions. Such second generation vectors were evaluated in vivo. These studies have revealed the complexity of the immune mechanisms elicited by these vectors and the importance of several parameters in these evaluations (i.e. mouse strains, nature of the transgene, route of administration...). In order to inhibit the production of neutralizing antibodies to adenovirus that prevent from further readministration of the vectors, immunosuppressive strategies were undertaken. Treatment regimens with immunosuppressive drugs (cyclophosphamide, FK506) or with monoclonal antibodies that block either the T cell receptor or costimulation pathways allow prolonged transgene expression and/or readministration of adenoviral vectors. In addition, transduction efficiencies may be increased by transiently inhibiting non-specific immune mechanisms that lead to the dramatic early clearance of the vectors. Taken together, these strategies may improve further gene therapy protocols by decreasing the host immune response to adenoviral vectors.


Subject(s)
Adenoviruses, Human/immunology , Genetic Therapy , Genetic Vectors/immunology , Adenoviruses, Human/genetics , Animals , Gene Expression , Humans , Mice , Neutralization Tests , Recombination, Genetic , T-Lymphocytes, Cytotoxic/immunology , Transgenes
14.
Plant Physiol ; 112(4): 1437-46, 1996 Dec.
Article in English | MEDLINE | ID: mdl-8972593

ABSTRACT

Primary expression of the Rhizobium meliloti-induced peroxidase gene rip1 occurs prior to nodule morphogenesis, specifically at the site of impending rhizobial infection (D. Cook, D. Dreyer, D. Bonnet, M. Howell, E. Nony, K. VandenBosch [1995] Plant Cell 7: 43-55). We examined the distribution and structure of rip1 transcript throughout nodule development. We determined that expression of rip1 in root tips is correlated with the competence of this zone for symbiotic association, whereas after rhizobial infection rip1 transcript is specifically associated with the zone of nodule development, including nascent nodule primordia. rip1 transcripts are characterized by multiple polyadenylation sites distributed within 200 to 400 bp of the translation stop site, and a single major transcription initiation site in close proximity to the rip1 open reading frame. Thus, rip1 expression is likely to be mediated through effects on a single transcription unit. Immediately 5' of the rip1 transcription unit DNA sequence analysis identified a 377-bp DNA element containing extensive repeat structure that is widely distributed in the Medicago truncatula genome.


Subject(s)
Gene Expression Regulation, Bacterial , Gene Expression Regulation, Enzymologic , Peroxidases/genetics , Sinorhizobium meliloti/genetics , Base Sequence , DNA Transposable Elements , DNA, Complementary , Molecular Sequence Data , Transcription, Genetic
15.
Gene Ther ; 3(1): 75-84, 1996 Jan.
Article in English | MEDLINE | ID: mdl-8929914

ABSTRACT

Replication-defective E1-deleted adenoviruses are attractive vectors for gene therapy or live vaccines. However, manufacturing methods required for their pharmaceutical development are not optimized. For example, the generation of E1-deleted adenovirus vectors relies on the complementation functions present in 293 cells. However, 293 cells are prone to the generation of replication competent particles as a result of recombination events between the viral DNA and the integrated adenovirus sequences present in the cell line. We report here that human lung A549 cells transformed with constitutive or inducible E1-expression vectors support the replication of E1-deficient adenoviruses. E1A transcription was elevated in most of the cell lines, and E1A proteins were expressed at levels similar to those of 293 cells. However, the levels of expression of E1A did not correlate with the efficiencies of complementation of E1-deleted viruses in A549 clones, since some clones complemented replication in the absence of induction of E1A expression. In addition, complementation of E1-deficient adenoviruses did not require expression of the E1B 55-kDa protein. Although these cell lines contain the coding and cis-acting regulatory sequences of the structural protein IX gene, they are not able to complement viruses in which this gene has been deleted. In contrast to 293 cells, such new complementation cell lines do not contain the left end of the adenoviral genome and thus represent a significant improvement over the currently used 293 cells, in which a single recombination event is sufficient to yield replication competent adenovirus.


Subject(s)
Adenoviruses, Human/genetics , Capsid Proteins , Carcinoma/metabolism , Gene Expression Regulation, Viral , Genetic Vectors/genetics , Lung Neoplasms/metabolism , Adenovirus E1 Proteins/deficiency , Adenovirus E1 Proteins/genetics , Adenovirus E1A Proteins/genetics , Adenovirus E1B Proteins/genetics , Adenoviruses, Human/growth & development , Animals , Capsid/genetics , Carcinoma/pathology , Chlorocebus aethiops , Gene Expression , Genetic Complementation Test , Humans , Lung Neoplasms/pathology , Plasmids , Transfection , Tumor Cells, Cultured , Vero Cells
16.
Brain Res Mol Brain Res ; 31(1-2): 210-4, 1995 Jul.
Article in English | MEDLINE | ID: mdl-7476031

ABSTRACT

Previously we reported that a single injection of nicotine decreased AP-1 DNA binding activity in adrenal medullae, although chronic bidaily nicotine (and saline) injections increased this binding activity [15]. Repeated acute nicotine injections (3 mg/kg i.p., 7 injections equi-spaced over a 3 h period) effectively increased adrenal tyrosine hydroxylase [3] and [Met5]enkephalin levels and also profoundly decreased adrenal medulla AP-1 DNA binding activity for over 8 h.


Subject(s)
Adrenal Medulla/drug effects , DNA-Binding Proteins/metabolism , Enkephalins/metabolism , Nicotine/administration & dosage , Transcription Factor AP-1/metabolism , Adrenal Medulla/metabolism , Animals , Base Sequence , Drug Administration Schedule , Female , Male , Molecular Sequence Data , Rats , Rats, Inbred F344 , Tyrosine 3-Monooxygenase/drug effects , Tyrosine 3-Monooxygenase/metabolism
17.
Gene Ther ; 2(4): 263-8, 1995 Jun.
Article in English | MEDLINE | ID: mdl-7552986

ABSTRACT

Adenoviruses are efficient gene transfer vectors for a variety of cell types. To date, the most widely used methods to construct recombinant adenoviruses involve either in vitro ligation or recombination between one-half of the virus genome, previously cloned in a plasmid vector and engineered to contain the desired expression cassette, and the other half of the virus genome prepared from virions. Although quite effective, these approaches produce viral progeny containing a mixture of recombinant and parental background virus. Thus the recovery of the recombinant virus can be difficult, especially when it grows more slowly than the parental virus. To improve selection and recovery of recombinant adenoviruses, we have constructed an adenovirus vector, AdTG6553, in which the E1 region has been replaced by the thymidine kinase (tk) gene of herpes simplex virus type 1. We show that infection of cells with AdTG6553 in the presence of the nucleoside analog ganciclovir (GCV) prevents viral replication. The conditional lethal phenotype introduced in AdTG6553 makes it a valuable tool to counter-select parental background virus in the presence of GCV and isolate replication-deficient recombinant adenoviruses in which the tk expression cassette has been replaced by a new gene.


Subject(s)
Adenoviridae/genetics , Genetic Vectors , Recombination, Genetic , Adenoviridae/drug effects , Adenoviridae/isolation & purification , Base Sequence , Ganciclovir/pharmacology , Gene Expression , Humans , Immediate-Early Proteins/biosynthesis , Immediate-Early Proteins/genetics , Molecular Sequence Data , Simplexvirus/drug effects , Simplexvirus/genetics , Simplexvirus/isolation & purification , Thymidine Kinase/biosynthesis , Thymidine Kinase/genetics , Tumor Cells, Cultured , Virus Cultivation/methods , Virus Replication/drug effects
18.
Hum Gene Ther ; 6(6): 711-21, 1995 Jun.
Article in English | MEDLINE | ID: mdl-7548271

ABSTRACT

Treatment of cystic fibrosis by gene therapy will require the development of vectors capable of efficient and safe transfer of a functional cystic fibrosis transmembrane conductance regulator (CFTR) cDNA to airway epithelia. To achieve this goal, replication-deficient (E1-) adenoviruses (Ad) are promising vectors. We have previously demonstrated efficient CFTR gene delivery to the airways of cotton rats and rhesus monkeys using a replication-deficient adenovirus, Ad-CFTR. Here, we have investigated an important safety issue, the interaction between the vector and wild-type virus which can provide the missing E1 function in trans. We show that Ad5 can mobilize the defective Ad-CFTR genome in vitro and in cotton rats. However, the extent of the complementation in vivo by wild-type virus is limited because no additional spreading or shedding of Ad-CFTR to trachea, lungs, and stools is elicited. To attenuate Ad-CFTR further, a mutation was introduced in the cis-acting regulatory sequences that control the encapsidation of the viral genome. We demonstrate that when cells are coinfected with wild-type virus and the new attenuated vector, the viral DNA containing the natural encapsidation sequences is preferentially packaged, leading to a rapid dilution of the recombinant virus.


Subject(s)
Adenoviridae/genetics , Adenovirus E1 Proteins/genetics , Cystic Fibrosis Transmembrane Conductance Regulator/genetics , Cystic Fibrosis/therapy , Genetic Therapy , Genetic Vectors , Animals , Base Sequence , Cell Line , DNA, Viral/genetics , Defective Viruses/genetics , Female , Genetic Complementation Test , Humans , Male , Molecular Sequence Data , Oligodeoxyribonucleotides , Sequence Deletion , Sigmodontinae
19.
Plant Cell ; 7(1): 43-55, 1995 Jan.
Article in English | MEDLINE | ID: mdl-7696879

ABSTRACT

Although key determinative events of the Rhizobium-legume symbiosis are likely to precede bacterial infection, no plant genes have been identified that are expressed strongly prior to infection and nodule morphogenesis. A subtractive hybridization-polymerase chain reaction technique was used to enrich for genes induced during the early phases of the R. meliloti-Medicago truncatula symbiosis. One gene so identified encodes a putative plant peroxidase protein, which we have named Rip1 for Rhizobium-induced peroxidase. The accumulation of rip1 transcript was rapidly and transiently induced by R. meliloti and by the corresponding lipooligosaccharide signal molecule Nod factor RmIV, which was both necessary and sufficient for rip1 induction. The duration of maximal rip1 expression coincided with the preinfection period: transcript levels for rip1 were near maximal by 3 hr postinoculation and declined by 48 hr, coincident with early infection events and the onset of nodule morphogenesis. Furthermore, although rip1 induction preceded bacterial infection by at least 24 hr, the transcript was localized to epidermal cells in the differentiating root zone that was subsequently infected by Rhizobium. Thus, a defining feature of the Rhizobium infection court is the prior induction of rip1 expression.


Subject(s)
Medicago sativa/genetics , Peroxidases/genetics , Sinorhizobium meliloti/pathogenicity , Amino Acid Sequence , Base Sequence , Cloning, Molecular , DNA, Complementary , Gene Expression Regulation, Enzymologic , Gene Expression Regulation, Plant , Kinetics , Medicago sativa/enzymology , Medicago sativa/microbiology , Molecular Sequence Data , Sequence Homology, Amino Acid
20.
Brain Res Mol Brain Res ; 19(4): 349-52, 1993 Sep.
Article in English | MEDLINE | ID: mdl-8231739

ABSTRACT

Nuclear extracts from hippocampi, striata and hypothalami of postnatal day (P) 7 rats contained elevated basal levels of AP-1 DNA binding activity and c-jun protein, which decreased to the low basal levels observed in the adult by P21. In contrast to the AP-1 DNA binding complex in the adult brain, the fos-related antigens were not a major component of the P7 AP-1 DNA binding activity.


Subject(s)
Aging/metabolism , Brain/metabolism , DNA-Binding Proteins/metabolism , Proto-Oncogene Proteins c-jun/metabolism , Animals , Base Sequence , Brain/growth & development , Cell Nucleus/metabolism , Corpus Striatum/growth & development , Corpus Striatum/metabolism , DNA-Binding Proteins/isolation & purification , Genes, jun , Hippocampus/growth & development , Hippocampus/metabolism , Hypothalamus/growth & development , Hypothalamus/metabolism , Male , Molecular Sequence Data , Oligonucleotide Probes , Proto-Oncogene Proteins c-jun/biosynthesis , Proto-Oncogene Proteins c-jun/isolation & purification , Rats , Rats, Inbred F344
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