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1.
Alzheimers Dement ; 16(1): 60-70, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31914226

RESUMO

INTRODUCTION: Behavioral variant frontotemporal dementia (bvFTD) may present sporadically or due to an autosomal dominant mutation. Characterization of both forms will improve understanding of the generalizability of assessments and treatments. METHODS: A total of 135 sporadic (s-bvFTD; mean age 63.3 years; 34% female) and 99 familial (f-bvFTD; mean age 59.9; 48% female) bvFTD participants were identified. f-bvFTD cases included 43 with known or presumed chromosome 9 open reading frame 72 (C9orf72) gene expansions, 28 with known or presumed microtubule-associated protein tau (MAPT) mutations, 14 with known progranulin (GRN) mutations, and 14 with a strong family history of FTD but no identified mutation. RESULTS: Participants with f-bvFTD were younger and had earlier age at onset. s-bvFTD had higher total Neuropsychiatric Inventory Questionnaire (NPI-Q) scores due to more frequent endorsement of depression and irritability. DISCUSSION: f-bvFTD and s-bvFTD cases are clinically similar, suggesting the generalizability of novel biomarkers, therapies, and clinical tools developed in either form to the other.


Assuntos
Demência Frontotemporal , Predisposição Genética para Doença , Mutação/genética , Testes Neuropsicológicos/estatística & dados numéricos , Fatores Etários , Idoso , Encéfalo/patologia , Proteína C9orf72/genética , Feminino , Demência Frontotemporal/classificação , Demência Frontotemporal/genética , Humanos , Masculino , Pessoa de Meia-Idade , América do Norte , Progranulinas/genética , Proteínas tau/genética
2.
Science ; 167(3920): 986-7, 1970 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-4188981

RESUMO

Daily business-day traffic determines the diurnal lead concentration as well as diurnal carbon monoxide concentration. Daily averages of 7.5 micrograms per cubic meter for lead and 13 parts per million of carbon monoxide were found for the 10-week period of the study. Correlations were demonstrated for lead and traffic and lead and carbon monoxide.


Assuntos
Poluição do Ar/análise , Monóxido de Carbono/análise , Chumbo/análise , Cidade de Nova Iorque , Periodicidade , Emissões de Veículos
3.
Science ; 182(4118): 1251-3, 1973 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-4796196

RESUMO

The helical wall-free microorganism, Spiroplasma citri, which is associated with citrus stubborn, a disease with no known vector, multiplied in the leafhopper vector of corn stunt but multiplied to higher titer in the vector of aster yellows and decreased the longevity of that insect. Acholeplasma laidlawii and A. granularum also multiplied in both leafhoppers.


Assuntos
Vetores de Doenças , Insetos , Mycoplasma/crescimento & desenvolvimento , Animais , Longevidade
4.
Science ; 182(4114): 827-9, 1973 Nov 23.
Artigo em Inglês | MEDLINE | ID: mdl-17772158

RESUMO

Growth-inhibition and precipitin tests established that antigens of the helical mycoplasma-like organism (Spiroplasma citri) associated with citrus stubborn disease are serologically related to antigens in corn infected with stunt disease but not in healthy corn.

5.
Plant Dis ; 93(3): 257-262, 2009 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30764183

RESUMO

In São Paulo State, Brazil, 'Candidatus Liberibacter americanus' and 'Candidatus Liberibacter asiaticus' are associated with huanglongbing (HLB). Affected municipalities occur mainly in the central and southern regions, where the annual number of hours above 30°C is two to five times lower than that in the extreme northern and western regions. The influence of temperature on sweet orange trees infected with 'Ca. L. asiaticus' or 'Ca. L. americanus' was studied in temperature-controlled growth chambers. Symptom progression on new shoots of naturally infected and experimentally graft-inoculated symptomatic sweet orange trees was assessed. Mottled leaves developed on all infected trees at 22 to 24°C, but not on any 'Ca. L. americanus'-infected trees at 27 to 32°C. Quantitative, real time-PCR was used to determine the liberibacter titers in the trees. After 90 days, 'Ca. L. asiaticus'-infected trees had high titers at 32 and 35°C, but not at 38°C, while 'Ca. L. americanus'-infected trees had high titers at 24°C, but at 32°C the titers were very low or the liberibacters could not be detected. Thus, the multiplication of 'Ca. L. asiaticus' is not yet affected at 35°C, while a temperature of 32°C is detrimental to 'Ca. L. americanus'. Thus, 'Ca. L. americanus' is less heat tolerant than 'Ca. L. asiaticus'. The uneven distribution of these two liberibacters in São Paulo State might be in relation with these results.

6.
Phytopathology ; 98(3): 337-44, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18944085

RESUMO

Brittle leaf disease or maladie des feuilles cassantes (MFC) is a lethal disorder of date palms that has assumed epidemic proportions in the oases of southern Tunisia. After a prolonged period during which palms are declining, the disease ends with the death of the palms. Whereas no pathogen could ever be associated with the disease, leaflets of affected palms have been previously shown to be deficient in manganese. Analysis of RNA preparations from leaflets of MFC-affected palms revealed the presence of a set of novel RNAs (MFC-RNAs) of sense and antisense polarities, which are homologous to various regions of the date palm chloroplast genome, such as the regions containing genes rrn5S-trnR(ACG) and trnM(CAU)-atpE. In the RNA preparations obtained from leaflets of affected palms, some of these RNAs are present as double-stranded species (MFC-dsRNAs), as witnessed by results from cellulose chromatography, end labeling, RNase digestion, and northern hybridization with strand specific probes. These MFC-RNAs represent a novel type of host-derived RNAs, and their presence in MFC-affected date palms is of diagnostic value.


Assuntos
Arecaceae/genética , Genoma de Cloroplastos/genética , Folhas de Planta/genética , RNA de Cloroplastos/genética , Arecaceae/virologia , Northern Blotting , Interações Hospedeiro-Patógeno , Doenças das Plantas/genética , Doenças das Plantas/virologia , Folhas de Planta/virologia , Vírus de Plantas/fisiologia , RNA de Cloroplastos/metabolismo
7.
Phytopathology ; 98(9): 977-84, 2008 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-18943735

RESUMO

In February 2007, sweet orange trees with characteristic symptoms of huanglongbing (HLB) were encountered in a region of São Paulo state (SPs) hitherto free of HLB. These trees tested negative for the three liberibacter species associated with HLB. A polymerase chain reaction (PCR) product from symptomatic fruit columella DNA amplifications with universal primers fD1/rP1 was cloned and sequenced. The corresponding agent was found to have highest 16S rDNA sequence identity (99%) with the pigeon pea witches'-broom phytoplasma of group 16Sr IX. Sequences of PCR products obtained with phytoplasma 16S rDNA primer pairs fU5/rU3, fU5/P7 confirm these results. With two primers D7f2/D7r2 designed based on the 16S rDNA sequence of the cloned DNA fragment, positive amplifications were obtained from more than one hundred samples including symptomatic fruits and blotchy mottle leaves. Samples positive for phytoplasmas were negative for liberibacters, except for four samples, which were positive for both the phytoplasma and 'Candidatus Liberibacter asiaticus'. The phytoplasma was detected by electron microscopy in the sieve tubes of midribs from symptomatic leaves. These results show that a phytoplasma of group IX is associated with citrus HLB symptoms in northern, central, and southern SPs. This phytoplasma has very probably been transmitted to citrus from an external source of inoculum, but the putative insect vector is not yet known.


Assuntos
Citrus/microbiologia , Phytoplasma/genética , Doenças das Plantas/microbiologia , RNA Ribossômico 16S/genética , Brasil , Clonagem Molecular , Primers do DNA , DNA Bacteriano/genética , DNA Ribossômico/genética , Phytoplasma/classificação , Phytoplasma/patogenicidade , Reação em Cadeia da Polimerase , RNA Bacteriano/genética
8.
Rev Chir Orthop Reparatrice Appar Mot ; 94(3): 252-60, 2008 May.
Artigo em Francês | MEDLINE | ID: mdl-18456060

RESUMO

PURPOSE OF THE STUDY: The increasing popularity of total-knee arthroplasty has led to many technical improvements both in the field of prosthesis design and implanted material and instrumentation. The recent advent of computer-assisted techniques is the fruit of a search for more precision for the bone cuts and better ligament balance. The purpose of the present study was to demonstrate how easy it is to use navigation systems by examining the difficulties encountered by one operator with navigation experience when the material was changed. MATERIAL AND METHODS: The first 30 total-knee arthroplasties implanted with a new navigation system were investigated. Elements specifically related to navigation difficulties were studied. The series was composed of 16 women and 14 men, mean age 65.9 years at the time of surgery (range, 43 to 84). Mean BMI was 30.66 (range, 23.05 to 39.54). All patients were reviewed by the operator using a standard X-ray protocol. Mean follow-up was six months. The 30 arthroplasties were consecutive, with no exclusions excepting revision procedures. Primary or post-traumatic degeneration was the main reason for surgery. This series was compared with two prior series of 30 prostheses each, implanted with a different navigation system. The first 30 and last 30 implantations using the previous navigation system were thus compared in terms of operative time and precision (comparison of postoperative alignment and implant position). The study focused on difficulties encountered when using the new system, on intra- and postoperative complications and on assessment of implant position. RESULTS: All procedures were totally performed with the navigation system, no interruptions. Operative time was lengthened by an average of 18 min (range, 0 to 45 min). There were no complications specifically related to the navigation system. The position of the implants was assessed in the frontal and sagittal plane on the plain X-rays and with a goniometer. Computed tomography was used to assess femoral component rotation. The overall alignment of the lower limb was within the "ideal" range of +/-3 degrees in 97% (average 0.1 degrees varus). The position of the femoral implant and the tibial plate was correct in the frontal and sagittal planes and no internal rotation of the femoral piece was noted on the 27 ct scan studies (mean 1.9 degrees external rotation). Implant accuracy was equivalent to that observed in the series of the last 30 implants using the prior navigation system. The learning curve was shorter. DISCUSSION: This small series demonstrated the absence of major problems with the new navigation system. The length of the learning curve was acceptable. This study demonstrated that prior experience with navigation is beneficial because the learning curve with the new system was shorter and the accuracy of implantation was equivalent to that achieved with the prior system. Widespread use of computer-assisted surgery should enable continued improvement in ancillary systems in the upcoming years, particularly concerning rotatory position of the femoral implants, which is still a problem. Cost containment will also be a necessary goal.


Assuntos
Artroplastia do Joelho/métodos , Competência Clínica , Cirurgia Assistida por Computador/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Artroplastia do Joelho/instrumentação , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Cirurgia Assistida por Computador/instrumentação
9.
Phytopathology ; 96(4): 356-68, 2006 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18943417

RESUMO

ABSTRACT Citrus exocortis viroid (CEVd), Citrus bent leaf viroid (CBLVd), a noncachexia variant of Hop stunt viroid (HSVd), Citrus viroid III (CVd-III), and Citrus viroid IV (CVd-IV) were co-inoculated as two-, three-, four-, and five-viroid mixtures to Clementine trees grafted on trifoliate orange to evaluate their effect on symptom expression, tree growth, and fruit yield. Most trees infected with CEVd-containing viroid mixtures developed exocortis scaling symptoms, as did CEVd alone, whereas most trees infected with HSVd- or CVd-IV-containing mixtures developed bark-cracking symptoms. Trees infected with mixtures containing both CEVd and CVd-IV revealed the existence of antagonism between these two viroids in terms of the expected bark-scaling and cracking symptoms. Synergistic interactions also were identified in trees infected with certain viroid combinations that, in spite of lacking CEVd, expressed exocortis-like scaling symptoms. Viroid interactions also affected the expected response of trees in terms of vegetative growth and fruit yield. Trees infected with viroid combinations containing CEVd or CVd-III were smaller and produced less fruit than trees infected with mixtures not containing these viroids. Viroid interactions on scion circumference and cumulative fruit yield, in terms of additivity of their effects, were statistically confirmed using a factorial analysis of variance model with two mean estimation approaches. In single-viroid infections, CEVd, CVd-III, and, to a lesser extent, CBLVd consistently and significantly reduced tree size and fruit yield. Conversely, HSVd and CVd-IV slightly increased fruit yield and reduced scion circumference. Rare and not consistent significant interactions were detected with the five-, four-, and three-viroid combinations. Antagonistic interactions between CEVd and CVd-III or CBLVd and CVd-III were revealed over the years with consistent significance. The antagonistic interaction between CEVd and CVd-IV was highly significant over the years when additional viroids were present; however, this antagonism appeared much later in the case of an exclusive interaction. HSVd and CVd-IV showed a consistent and significant synergistic interaction on yield only when both viroids were exclusively present. These results demonstrate antagonistic or synergistic relationships between citrus viroids depending on the viroid mixtures present in the host.

10.
Arch Intern Med ; 139(3): 317-20, 1979 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-426576

RESUMO

Thirty-eight episodes of culture-documented antibiotic-resistant bacterial or fungal infection in patients with malignant neoplasms were treated with daily granulocyte transfusions until the infection improved or the patient died. Cumulative summation temperature plotting allowed easier interpretation of recipient fever response. Seventy-one percent of recipients had a favorable response to transfusion. There was no difference in mortality between patients treated with cells collected by filtration (FL) or intermittent flow centrifugation (IFCL) leukapheresis techniques. Transfusion reactions were more than twice as common with FL than IFCL collected cells. Seventy-four percent of recipients were alive 21 days after completion of transfusions; of the ten deaths, five could be classified as granulocyte transfusion failures. This study suggests that long-term granulocyte transfusion may be required in infected recipients when autologous granulocytes do not return after chemotherapy.


Assuntos
Transfusão de Sangue , Granulócitos , Neoplasias/complicações , Sepse/terapia , Abscesso/etiologia , Abscesso/terapia , Adulto , Celulite (Flegmão)/etiologia , Celulite (Flegmão)/terapia , Humanos , Pneumonia/etiologia , Pneumonia/terapia , Doenças Retais/etiologia , Doenças Retais/terapia , Sepse/etiologia , Fatores de Tempo
11.
Plant Dis ; 89(10): 1129, 2005 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30791288

RESUMO

In October 2003, during a survey to evaluate the incidence of phytoplasma diseases in Lebanon, symptoms suggestive of phytoplasma infection in Opuntia monacantha (Haworth) were observed in Saghbine, Bekaa Valley. Symptoms were excessive stem and shoot proliferation. Three symptomatic and as well as symptomless plants were collected and analyzed for the presence of phytoplasmas. Nucleic acids were extracted from 0.5 g of shoot tissue and tested using polymerase chain reaction (PCR) with universal phytoplasma primers (fU5rU3) for partial amplification of the ribosomal 16SrDNA (4). PCR resulted in amplification of an expected 881-bp rDNA fragment from the symptomatic but not from symptomless samples. For characterization, sequence of the amplified DNA was determined (Genbank Accession No. AY939815). The sequence showed a high similarity with several isolates of the 16srII group of phytoplasmas. The highest similarity has been oserved with 16S rDNA of two isolates of cactus witches'-broom phytoplasma found in China (1) and Mexico (3) (Genbank Accession Nos. AJ293216 and AF320575, respectively) (99.8%) as well as faba bean phyllody phytoplasma (Genbank Accession No. X83432) (99.7%) and "Candidatus Phytoplasma aurantifolia" (Genbank Accession No. U15442) (99.3%). The presence of phytoplasmas was confirmed using nested-PCR with primers R16mF2/R1 and R16F2n/R2 (2). The Tru9I digestion pattern of the amplified product R16F2n/F16R2 detected in O. monacantha was identical to the digestion pattern obtained from periwinkle infected by "Ca. P. aurantifolia" (subgroup 16SrII-B) and soybean phyllody phytoplasma (subgroup 16SrII-C), but different from the Tru9I digestion pattern observed for cleome phyllody phytoplasma (subgroup 16SrII-A) and tomato big bud phytoplasma (subgroup 16SrII-E). To our knowledge, this is the first report of an infection with a phytoplasma belonging to16SrII group in Lebanon. References: (1) H. Cai et al. Plant Pathol. 51:394, 2002. (2) D. E. Gundersen and I. M. Lee. Phytopathol. Mediterr. 35:144, 1996. (3) N. E. Leyva-Lopez et al. Phytopathology. (Abstr.) 89(suppl):S45, 1999. (4) B. Schneider et al. Pages 369-380 in: Molecular and Diagnostic Procedures in Mycoplasmology. Academic Press, NY, 1995.

12.
Plant Dis ; 89(1): 107, 2005 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30795297

RESUMO

Huanglongbing (HLB) (ex-greening) is one of the most serious diseases of citrus. The causal agent is a noncultured, sieve tube-restricted α-proteobacterium, "Candidatus Liberibacter africanus" in Africa and "Candidatus Liberibacter asiaticus" in Asia (2). The disease has never been reported from the American continent. However, Diaphorina citri, the Asian psyllid vector of HLB, is found in South, Central, and North America (Florida and Texas). Early in 2004, leaf and fruit symptoms resembling those of HLB were observed in several sweet orange orchards near the city of Araraquara, Sao Paulo State. Leaf mottling on small and large leaves was the major symptom. Shoots with affected leaves were yellowish. Fruits were small and lopsided, contained many aborted seeds, and appeared more severely affected than were plants infected with classic HLB. Forty-three symptomatic samples and twenty-five samples of symptomless sweet orange leaves from five farms were analyzed for the presence of the HLB-liberibacters using polymerase chain reaction (PCR) with two sets of HLB-specific primers for amplification of 16S rDNA (2,3) and ribosomal protein genes (1). None of the 43 symptomatic leaf samples gave a positive PCR amplification, while HLB-affected leaves from the Bordeaux HLB collection produced the characteristic amplicons with both sets of primers. The 43 symptomatic and the 25 symptomless leaf samples were then analyzed using PCR with universal primers for amplification of bacterial 16S rDNA (4). All symptomatic leaf samples, but none of the symptomless leaf samples, yielded the same 16S rDNA amplification product, indicating the presence of a bacterium in the symptomatic leaves. This was confirmed using the observation of a sieve tube restricted bacterium by electron microscopy. The 16S rDNA product was cloned, sequenced, and compared with those of "Ca. L. africanus" and "Ca. L. asiaticus". While the 16S rDNAs of these two liberibacter species have 97.5% sequence identity, the 16S rDNA sequence of the new bacterium shared only 93.7% identity with that of "Ca. L. asiaticus" and 93.9% with that of "Ca. L. africanus". The 16S rDNA sequence of the new bacterium had a secondary loop structure characteristic of the α subdivision of the proteobacteria and possessed all the oligonucleotide signatures characteristic of the liberibacters. For these reasons, the new bacterium is a liberibacter and is sufficiently different phylogenetically from known liberibacters to warrant a new species, "Candidatus Liberibacter americanus". Specific PCR primers for amplification of the 16S rDNA of the new species have been developed. They were able to detect "Ca. L. americanus" in 214 symptomatic leaf samples from 47 citrus farms in 35 municipalities, while the "old" species, "Ca. L. asiaticus", has been found only four times within the 47 farms. References: (1) A. Hocquellet et al. Mol. Cell. Probes, 13:373, 1999. (2) S. Jagoueix et al. Int. J. Syst. Bacteriol. 44:379, 1994. (3) S. Jagoueix et al. Mol. Cell. Probes 10:43, 1996. (4) W. G. Weisburg et al. J. Bacteriol. 173:697, 1991.

13.
Mol Plant Microbe Interact ; 13(10): 1145-55, 2000 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-11043476

RESUMO

Spiroplasma citri is a plant-pathogenic mollicute. Recently, the so-called nonphytopathogenic S. citri mutant GMT 553 was obtained by insertion of transposon Tn4001 into the first gene of the fructose operon. Additional fructose operon mutants were produced either by gene disruption or selection of spontaneous xylitol-resistant strains. The behavior of these spiroplasma mutants in the periwinkle plants has been studied. Plants infected via leafhoppers with the wild-type strain GII-3 began to show symptoms during the first week following the insect-transmission period, and the symptoms rapidly became severe. With the fructose operon mutants, symptoms appeared only during the fourth week and remained mild, except when reversion to a fructose+ phenotype occurred. In this case, the fructose+ revertants quickly overtook the fructose- mutants and the symptoms soon became severe. When mutant GMT 553 was complemented with the fructose operon genes that restore fructose utilization, severe pathogenicity, similar to that of the wild-type strain, was also restored. Finally, plants infected with the wild-type strain and grown at 23 degrees C instead of 30 degrees C showed late symptoms, but these rapidly became severe. These results are discussed in light of the role of fructose in plants. Fructose utilization by the spiroplasmas could impair sucrose loading into the sieve tubes by the companion cells and result in accumulation of carbohydrates in source leaves and depletion of carbon sources in sink tissues.


Assuntos
Frutose/metabolismo , Magnoliopsida/microbiologia , Óperon , Doenças das Plantas/microbiologia , Spiroplasma/metabolismo , Spiroplasma/patogenicidade , Animais , Genes Bacterianos , Teste de Complementação Genética , Glucose/metabolismo , Hemípteros/microbiologia , Mutagênese Insercional , Fenótipo , Spiroplasma/genética , Spiroplasma/crescimento & desenvolvimento , Xilitol/farmacologia
14.
Mol Plant Microbe Interact ; 14(2): 225-33, 2001 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-11204786

RESUMO

A differential display of mRNAs was used to isolate periwinkle cDNAs differentially expressed following infection with one of three mollicutes: Spiroplasma citri, Candidatus Phytoplasma aurantifolia, and stolbur phytoplasma. Twenty-four differentially expressed cDNAs were characterized by Northern blots and sequence analysis. Eight of them had homologies with genes in databanks coding for proteins involved in photosynthesis, sugar transport, response to stress, or pathways of phytosterol synthesis. The regulation of these genes in periwinkle plants infected by additional phloem-restricted bacteria showed that they were not specific to a given mollicute, but correlations with particular symptoms could be established. Expression of transketolase was down regulated following infection with a pathogenic strain of S. citri. No down regulation was observed for the nonphytopathogenic mutant GMT553, which is deficient for fructose utilization.


Assuntos
Regulação Fúngica da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Magnoliopsida/genética , Mycoplasma/genética , Sequência de Aminoácidos , Sequência de Bases , DNA Complementar , Dados de Sequência Molecular , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Homologia de Sequência de Aminoácidos , Transcetolase/genética
15.
Gene ; 171(1): 95-8, 1996 May 24.
Artigo em Inglês | MEDLINE | ID: mdl-8675039

RESUMO

A 6.5-kb DNA fragment containing the gene (rpoB) encoding the RNA polymerase (RNAP) beta subunit, from the mollicute Spiroplasma citri (Sc), was cloned and sequenced. The classical eubacterial organization, with the genes (rplK, A, J and L) encoding ribosomal proteins L11, L1, L10 and L12 located immediately upstream from rpoB, was not found in the Sc DNA. Instead, an open reading frame (hsdS) potentially encoding a component of a type I restriction and modification system was identified upstream from rpoB, and sequences showing similarities with insertion elements were found between hsdS and rpoB.


Assuntos
Enzimas de Restrição-Modificação do DNA/genética , RNA Polimerases Dirigidas por DNA/genética , Genes Bacterianos/genética , Spiroplasma/genética , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , Desoxirribonucleases de Sítio Específico do Tipo I/genética , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Spiroplasma/enzimologia
16.
Gene ; 252(1-2): 61-9, 2000 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-10903438

RESUMO

Transposon Tn4001 mutagenesis of Spiroplasma citri wild-type (wt) strain GII-3 led to the isolation and characterization of non-phytopathogenic mutant GMT 553. In this mutant, transposon Tn4001 is inserted within the first gene of the fructose operon. This operon comprises three genes. The first gene (fruR) codes for a putative transcriptional regulator protein belonging to the deoxyribonucleoside repressor (DeoR) family. Sequence similarities and functional complementation of mutant GMT 553 with different combinations of the wt genes of the fructose operon showed that the second gene (fruA) codes for the permease of the phosphoenolpyruvate:fructose phosphotransferase system (fructose PTS), and the third, fruK, for the 1-phosphofructokinase (1-PFK). Transcription of the fructose operon in wt strain GII-3 resulted in two messenger RNAs, one of 2.8kb and one of 3.8kb. Insertion of Tn4001 in the genome of mutant GMT 553 abolished transcription of the fructose operon, and resulted in the inability of this mutant to use fructose. Functional complementation experiments demonstrated that fructose utilization was restored with fruR-fruA-fruK, fruA-fruK or fruA only, but not with fruR or fruR-fruA. This is the first time that an operon for sugar utilization has been functionally characterized in the mollicutes.


Assuntos
Frutose/metabolismo , Óperon/genética , Spiroplasma/metabolismo , Sequência de Aminoácidos , Proteínas de Bactérias/genética , Sequência de Bases , Sítios de Ligação , Elementos de DNA Transponíveis , DNA Bacteriano/química , DNA Bacteriano/genética , Teste de Complementação Genética , Proteínas de Membrana Transportadoras/genética , Dados de Sequência Molecular , Proteínas de Transporte de Monossacarídeos , Mutagênese Insercional , Mutação , Fatores de Iniciação de Peptídeos/genética , Fenótipo , Fosfofrutoquinase-1/genética , Mapeamento Físico do Cromossomo , Fator de Iniciação 2 em Procariotos , RNA Bacteriano/genética , RNA Bacteriano/metabolismo , Proteínas Repressoras/genética , Análise de Sequência de DNA , Spiroplasma/genética , Spiroplasma/patogenicidade , Fatores de Transcrição/genética , Transcrição Gênica
17.
Biochimie ; 75(8): 675-9, 1993.
Artigo em Inglês | MEDLINE | ID: mdl-8286440

RESUMO

In order to confirm the presence of a phosphoenolpyruvate (PEP)-dependent sugar-phosphotransferase system (PTS) in the mollicute Acholeplasma florum we studied the ability of cell free extracts of this organism to phosphorylate glucose and/or fructose in the presence of PEP. We also cloned and sequenced a DNA fragment coding for a putative polypeptide showing significant similarity with the enzyme II of the beta-glucoside PTS of Escherichia coli. Taken together, these results show that A florum possesses a fructose-PTS, but not a glucose-PTS, and that the amino acid sequence deduced from the DNA fragment is related to beta-glucoside and sucrose enzymes II of PTS from various bacteria.


Assuntos
Acholeplasma/enzimologia , Sistema Fosfotransferase de Açúcar do Fosfoenolpiruvato/metabolismo , Fosfotransferases (Aceptor do Grupo Nitrogenado)/metabolismo , Sequência de Aminoácidos , Clonagem Molecular , Escherichia coli/genética , Frutose/metabolismo , Glucose/metabolismo , Dados de Sequência Molecular , Sistema Fosfotransferase de Açúcar do Fosfoenolpiruvato/genética , Fosforilação , Fosfotransferases (Aceptor do Grupo Nitrogenado)/genética , Homologia de Sequência de Aminoácidos
18.
FEMS Microbiol Lett ; 198(1): 73-8, 2001 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-11325556

RESUMO

The role of fruR, the first gene of the Spiroplasma citri fructose operon, was investigated. In vivo transcription of the fructose operon is greatly enhanced by the presence of fructose in the growth medium while glucose has no effect. When fruR is not expressed, transcription of the fructose operon is not stimulated by fructose, and fructose fermentation is decreased, indicating that FruR is an activator of the fructose operon. The promoter of the fructose operon was localized by primer extension, and a direct T-rich repeat was found to overlap the -35 box. This repeat could be the binding site of FruR. The presence of fructose in the culture medium also decreases the toxicity of methyl alpha-glucoside, however FruR is not involved in this regulation. This is the first description of transcription regulation of a mollicute operon.


Assuntos
Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Frutose/metabolismo , Óperon , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Spiroplasma/genética , Transcrição Gênica , Sequência de Bases , Sítios de Ligação , Meios de Cultura , Fermentação , Regulação Bacteriana da Expressão Gênica , Teste de Complementação Genética , Glucose/metabolismo , Metilglucosídeos/farmacologia , Dados de Sequência Molecular , Regiões Promotoras Genéticas , Spiroplasma/metabolismo
19.
Vet Microbiol ; 16(4): 385-90, 1988 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-3381492

RESUMO

Mycoplasma salivarium, a common human oropharyngeal mycoplasma, was isolated from nasal and pharyngeal secretions of 14 of 284 swine in a barrier-maintained, disease-free herd. M. salivarium was recovered from one boar 6 times over a 26-month period and one time only from 13 other swine. Human isolates of M. salivarium were compared with the swine isolates by DNA-DNA hybridization and SDS-PAGE of the cell proteins and the strains were shown to be closely related. One of eight of the swine from which M. salivarium was isolated had complement-fixing antibodies and another culture-positive animal had metabolic-inhibiting antibodies to M. salivarium. Overt disease was not associated with the organism. These results support previous findings that mycoplasmas closely related to M. salivarium may be isolated from the nasopharynges of swine and they further indicate that the organism can establish persistently in swine without evidence of overt disease.


Assuntos
Infecções por Mycoplasma/veterinária , Mycoplasma/isolamento & purificação , Doenças dos Suínos/microbiologia , Animais , Proteínas de Bactérias/análise , DNA Bacteriano/análise , Eletroforese em Gel de Poliacrilamida , Humanos , Mycoplasma/classificação , Mycoplasma/genética , Infecções por Mycoplasma/microbiologia , Cavidade Nasal/microbiologia , Hibridização de Ácido Nucleico , Faringe/microbiologia , Suínos
20.
Vet Microbiol ; 76(1): 31-40, 2000 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-10925039

RESUMO

A nested polymerase chain reaction (PCR) was developed for the detection of Mycoplasma hyopneumoniae, the etiological agent of enzootic pneumonia, in tracheobronchiolar washings from live pigs. Two nested pairs of oligonucleotide primers were designed from the sequence of a specific DNA probe (I 141; accession number U02537). The primer combination was Hp1/Hp3 for the first step PCR while the nested primers (Hp4/Hp6) allowed amplification of a 706 bp fragment. All strains of M. hyopneumoniae tested in this study could be detected by the nested PCR. DNA from other bacterial species isolated from the respiratory tract of pigs or from other mycoplasmal species were not amplified. The detection limit was estimated to be 1 fg, corresponding approximately to one organism, while in the one step PCR previously described 4 x 10(2) organisms were required. The nested PCR was evaluated on 362 tracheobronchiolar lavages collected from pigs at 2, 4 and 6 months of age in eight herds chronically infected with M. hyopneumoniae. The nested PCR was compared to a blocking ELISA performed with sera collected from the same pigs at the same ages, and to an immunofluorescence test at slaughter on 65 lungs from 6-month old pigs. The comparison indicated that the nested PCR was significantly (p<0.05) more sensitive (157 positive results of 362 samples) than ELISA (118 positive results of 362 samples) for detection of M. hyopneumoniae infection. Nested PCR was also significantly more sensitive (54 positive results of 65 samples) than immunofluorescence (29 positive results of 65 samples) for detection of M. hyopneumoniae in pig lungs at slaughter. Moreover, the nested PCR was used to confirm the absence of the mollicute in a pig herd without any history of M. hyopneumoniae infection. Thus, nested PCR appears to be a useful test to assess M. hyopneumoniae infection on pig farms.


Assuntos
Infecções por Mycoplasma/veterinária , Mycoplasma/isolamento & purificação , Reação em Cadeia da Polimerase/veterinária , Doenças dos Suínos/diagnóstico , Animais , Brônquios/microbiologia , Ensaio de Imunoadsorção Enzimática/veterinária , Imunofluorescência/veterinária , Infecções por Mycoplasma/microbiologia , Reação em Cadeia da Polimerase/métodos , Sensibilidade e Especificidade , Suínos , Doenças dos Suínos/microbiologia , Traqueia/microbiologia
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