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1.
Mol Ecol ; 33(5): e17268, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38230514

RESUMEN

Ecological divergence due to habitat difference plays a prominent role in the formation of new species, but the genetic architecture during ecological speciation and the mechanism underlying phenotypic divergence remain less understood. Two wild ancestors of rice (Oryza rufipogon and Oryza nivara) are a progenitor-derivative species pair with ecological divergence and provide a unique system for studying ecological adaptation/speciation. Here, we constructed a high-resolution linkage map and conducted a quantitative trait locus (QTL) analysis of 19 phenotypic traits using an F2 population generated from a cross between the two Oryza species. We identified 113 QTLs associated with interspecific divergence of 16 quantitative traits, with effect sizes ranging from 1.61% to 34.1% in terms of the percentage of variation explained (PVE). The distribution of effect sizes of QTLs followed a negative exponential, suggesting that a few genes of large effect and many genes of small effect were responsible for the phenotypic divergence. We observed 18 clusters of QTLs (QTL hotspots) on 11 chromosomes, significantly more than that expected by chance, demonstrating the importance of coinheritance of loci/genes in ecological adaptation/speciation. Analysis of effect direction and v-test statistics revealed that interspecific differentiation of most traits was driven by divergent natural selection, supporting the argument that ecological adaptation/speciation would proceed rapidly under coordinated selection on multiple traits. Our findings provide new insights into the understanding of genetic architecture of ecological adaptation and speciation in plants and help effective manipulation of specific genes or gene cluster in rice breeding.


Asunto(s)
Oryza , Oryza/genética , Fitomejoramiento , Mapeo Cromosómico , Fenotipo , Sitios de Carácter Cuantitativo/genética
2.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 46(2): 297-300, 2024 Apr.
Artículo en Zh | MEDLINE | ID: mdl-38686729

RESUMEN

Bone infarction has a low incidence in clinical practice and mostly occurs in the metaphysis and diaphysis.Few studies report the advanced imaging technique for bone infarction.Here we reported the fast field echo resembling a CT using restricted echo-spacing and calcium-suppressed spectral CT imaging for a case of multifocal bone infarcts in both lower extremities,aiming to provide diagnostic experience for clinical practice.


Asunto(s)
Infarto , Imagen por Resonancia Magnética , Tomografía Computarizada por Rayos X , Humanos , Masculino , Calcio , Infarto/diagnóstico por imagen , Extremidad Inferior/diagnóstico por imagen , Extremidad Inferior/irrigación sanguínea , Imagen por Resonancia Magnética/métodos , Tomografía Computarizada por Rayos X/métodos , Adulto
3.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 44(4): 733-736, 2022 Aug.
Artículo en Zh | MEDLINE | ID: mdl-36065709

RESUMEN

The diagnosis of atypical cystic meningioma is difficult because of the atypical MRI findings.This paper reported a case of atypical cystic meningioma and described the detailed MRI findings,which would provide valuable imaging evidence for the localization and classification of cystic meningioma.


Asunto(s)
Neoplasias Meníngeas , Meningioma , Humanos , Imagen por Resonancia Magnética , Neoplasias Meníngeas/diagnóstico por imagen , Meningioma/diagnóstico por imagen
4.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 43(2): 300-304, 2021 Apr 28.
Artículo en Zh | MEDLINE | ID: mdl-33966714

RESUMEN

Intracranial intradural chondroma is a rare disorder,the imaging findings of which have been rarely reported.The current study reported a case of intracranial extra-cerebral chondroma and described the detailed CT and magnetic resonance imaging findings,which would provide valuable imaging evidence for the diagnosis of intracranial extra-cerebral chondroma.


Asunto(s)
Neoplasias Encefálicas , Condroma , Neoplasias Encefálicas/diagnóstico por imagen , Condroma/diagnóstico por imagen , Humanos , Imagen por Resonancia Magnética , Tomografía Computarizada por Rayos X
5.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 43(1): 57-62, 2021 Feb 28.
Artículo en Zh | MEDLINE | ID: mdl-33663664

RESUMEN

Objective To investigate the intra-and inter-observer reproducibility of iodine concentrations of abdominal parenchymal organs based on spectral CT.Methods The water-free iodine images of the venous phase were retrospectively obtained from 50 patients with abdominal dynamic spectral CT scans.The iodine concentrations were measured in the left,right and caudate lobes of liver,spleen,pancreas and bilateral kidneys.Intraclass correlation coefficient(ICC)and Bland-Altman plot were employed to analyze the intra-and inter-observer reproducibility.Results The intra-observer ICCs of the left,right and caudate lobes of liver,spleen,pancreas,and left and right kidneys were 0.938(0.894,0.965),0.932(0.884,0.961),0.939(0.895,0.965),0.947(0.909,0.970),0.912(0.851,0.949),0.946(0.906,0.969)and 0.907(0.842,0.946),which indicated good intra-observer reproducibility.The inter-observer ICCs of the left,right and caudate lobes of liver,spleen,pancreas,and left and right kidneys were 0.947(0.909,0.970),0.927(0.875,0.958),0.943(0.902,0.968),0.956(0.924,0.975),0.934(0.887,0.962),0.927(0.875,0.958)and 0.892(0.818,0.937),which indicated good inter-observer reproducibility.Bland-Altman plots presented that more than 95% points of the intra-observer differences located within 95% CI of limits of agreement for the caudate lobe of liver,spleen,pancreas and bilateral kidneys,which was same as inter-observer differences of the caudate lobe of liver,spleen and right kidney.Conclusion The iodine concentration measurement based on the spectral CT presented good intra-and inter-observer reproducibility for the caudate lobe of liver and spleen.


Asunto(s)
Yodo , Humanos , Variaciones Dependientes del Observador , Reproducibilidad de los Resultados , Estudios Retrospectivos , Tomografía Computarizada por Rayos X
6.
Mol Biol Evol ; 36(5): 875-889, 2019 05 01.
Artículo en Inglés | MEDLINE | ID: mdl-30861529

RESUMEN

The occurrence of parallel speciation strongly implies the action of natural selection. However, it is unclear how general a phenomena parallel speciation is since it was only shown in a small number of animal species. In particular, the adaptive process and mechanisms underlying the process of parallel speciation remain elusive. Here, we used an integrative approach incorporating population genomics, common garden, and crossing experiments to investigate parallel speciation of the wild rice species Oryza nivara from O. rufipogon. We demonstrated that O. nivara originated multiple times from different O. rufipogon populations and revealed that different O. nivara populations have evolved similar phenotypes under divergent selection, a reflection of recurrent local adaptation of ancient O. rufipogon populations to dry habitats. Almost completed premating isolation was detected between O. nivara and O. rufipogon in the absence of any postmating barriers between and within these species. These results suggest that flowering time is a "magic" trait that contributes to both local adaptation and reproductive isolation in the origin of wild rice species. Our study thus demonstrates a convincing case of parallel ecological speciation as a consequence of adaptation to new environments.


Asunto(s)
Especiación Genética , Oryza/genética , Adaptación Biológica , Asia Sudoriental , Asia Occidental , Ecosistema , Fenotipo , Filogeografía , Polimorfismo de Nucleótido Simple , Aislamiento Reproductivo , Selección Genética , Secuenciación Completa del Genoma
7.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 42(6): 776-780, 2020 Dec 30.
Artículo en Zh | MEDLINE | ID: mdl-33423725

RESUMEN

Objective To identify the optimal mono-energetic enhanced spectral CT for renal cortex in cortical phase based on the iodine concentration. Methods Fifty patients with normal renal function received the abdominal enhanced spectral CT examination.The iodine concentration and CT values of the multiple mono-energetic spectral images were measured on renal cortex in cortical phase,and the correlation between the iodine concentration and the CT values and the coefficient of variation(CV)were analyzed. Results The correlation analysis demonstrated that the correlation coefficient was 0.994,0.994,0.993,0.987,0.976,0.960,and 0.938 between mono-energetic spectral CT images(40-100 keV with interval 10 keV,respectively)and iodine concentration(all P <0.001).The correlation coefficients of 40 keV,50 keV(P=0.007)and 60 keV(P=0.030)were significantly higher compared with that of 70 keV,and the correlation coefficients were not significantly among 40 keV,50 keV and 60 keV(P =0.590).The CV of renal cortex in cortical phase was 0.21,0.20,and 0.19 for 40 keV,50 keV,and 60 keV,respectively. Conclusion 60 keV is the optimal mono-energy for the enhanced spectral CT of the renal cortex in cortical phase.


Asunto(s)
Medios de Contraste , Yodo , Corteza Renal/diagnóstico por imagen , Tomografía Computarizada por Rayos X , Humanos , Interpretación de Imagen Radiográfica Asistida por Computador
8.
Biotechnol Appl Biochem ; 64(4): 549-554, 2017 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-27301870

RESUMEN

Gene doping can be easily concealed since its product is similar to endogenous protein, making its effective detection very challenging. In this study, we selected insulin-like growth factor I (IGF-I) exogenous gene for gene doping detection. First, the synthetic IGF-I gene was subcloned to recombinant adeno-associated virus (rAAV) plasmid to produce recombinant rAAV2/IGF-I-GFP vectors. Second, in an animal model, rAAV2/IGF-I-GFP vectors were injected into the thigh muscle tissue of mice, and then muscle and blood specimens were sampled at different time points for total DNA isolation. Finally, real-time quantitative PCR was employed to detect the exogenous gene doping of IGF-I. In view of the characteristics of endogenous IGF-I gene sequences, a TaqMan probe was designed at the junction of exons 2 and 3 of IGF-I gene to distinguish it from the exogenous IGF-I gene. In addition, an internal reference control plasmid and its probe were used in PCR to rule out false-positive results through comparison of their threshold cycle (Ct) values. Thus, an accurate exogenous IGF-I gene detection approach was developed in this study.


Asunto(s)
Factor I del Crecimiento Similar a la Insulina/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Animales , Ensayo de Inmunoadsorción Enzimática , Vectores Genéticos/genética , Humanos , Ratones , Ratones Endogámicos ICR
9.
Mol Phylogenet Evol ; 85: 238-46, 2015 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-25732070

RESUMEN

Most plant phylogeographic studies in subtropical China have stressed the importance of multiple refugia and limited admixture among refugia. Little attention has been paid to range expansion and recolonization routes in this region. In this study, we implemented a phylogeographic survey on Sargentodoxa cuneata, a widespread woody deciduous climber in subtropical China to determine if it conforms to the expansion-contraction (EC) model during the Pleistocene. Sequence variation of two chloroplast intergenic spacers (IGSs) in 369 individuals from 54 populations of S. cuneata was examined. Twenty-six chloroplast haplotypes were recovered. One of these (H5) occurred across the range of S. cuneata and was absent from only 13 populations. Sixteen of the 26 haplotypes were connected to H5 by one mutation and displayed a star-like pattern in the haplotype network. All chloroplast haplotypes clustered into two lineages (A and B) in a Bayesian tree, and most haplotypes (18 out of 26) originated during the mid-Pleistocene (0.63-1.07Ma). Demographic analyses detected a recent range expansion that occurred at 95.98ka (CI: 61.7-112.53ka) for Lineage A. The genetic signature of an ancient range expansion after the Middle Pleistocene Transition (MPT) was also evident. Three recolonization routes were identified in subtropical China. The results suggest that temperate plants in subtropical China may conform to the EC model to some extent. However, the genetic signature from multiple historical processes may complicate the phylogeographic patterns of organisms in the region due to the mild Pleistocene climate. This study provides a new perspective for understanding the evolutionary history of temperate plants in subtropical China.


Asunto(s)
ADN de Cloroplastos/genética , Magnoliopsida/clasificación , Filogenia , Teorema de Bayes , Evolución Biológica , China , ADN de Plantas/genética , ADN Espaciador Ribosómico/genética , Variación Genética , Haplotipos , Magnoliopsida/genética , Filogeografía , Análisis de Secuencia de ADN
10.
BMC Microbiol ; 14: 37, 2014 Feb 12.
Artículo en Inglés | MEDLINE | ID: mdl-24521422

RESUMEN

BACKGROUND: Latent tuberculosis infection (LTBI) relies on a homeostasis of macrophages and Mycobacterium tuberculosis (Mtb). The small heat shock protein, Mtb Hsp16.3 (also known as latency-associated antigen), plays an important role in Mtb persistence within macrophages. However, the mechanism of LTBI remains elusive. The aim of this study was to delineate LTBI-related miRNA expression in U937 macrophages expressing Mtb Hsp16.3 protein. U937 macrophages were infected with an integrase-deficient Lentivirus vector to transiently express Mtb Hsp16.3, and green fluorescent protein (GFP) as a control. We used a microRNA (miRNA) microarray chip containing more than 1000 probes to identify the significant differentially expressed miRNAs in the infected U937 cells, and employed real-time quantitative polymerase chain reaction (qRT-PCR) for validation. Furthermore, we confirmed these candidate LTBI-related miRNAs in peripheral blood mononuclear cells from subjects with LTBI and in healthy control individuals. Functional annotation prediction of miRNA target genes and pathway enrichment analyses were used to explore the putative links between these miRNAs and LTBI. RESULTS: Analysis of the miRNA expression profile identified 149 miRNAs that were differentially expressed in U937 macrophages expressing Mtb Hsp16.3 compared with the control expressing GFP. The expression level of seven miRNAs (miR-424-5p, miR-493-5p, miR-296-5p, miR-27b-3p, miR-377-5p, miR-3680-5p, miR-191-5p) were validated by qRT-PCR. The expression level of four miRNAs (miR-424-5p, miR-27b-3p, miR-377-5p, miR-3680-5p) in the peripheral blood mononuclear cells samples from LTBI and healthy participants reflected the altered patterns observed in the microarray profile. The bioinformatic analyses suggest that the miRNAs may regulate Mtb latent infection by affecting the development of macrophage cells. CONCLUSIONS: The results suggest that miRNA expression may play a considerable role in the pathogenesis of LTBI, and this would increase our understanding of the molecular basis of Hsp16.3-facilitated Mtb survival in macrophages.


Asunto(s)
Proteínas Bacterianas/biosíntesis , Chaperoninas/biosíntesis , Interacciones Huésped-Patógeno , Tuberculosis Latente/inmunología , Tuberculosis Latente/microbiología , Macrófagos/microbiología , MicroARNs/biosíntesis , Mycobacterium tuberculosis/inmunología , Proteínas Bacterianas/genética , Sangre/inmunología , Células Cultivadas , Chaperoninas/genética , Perfilación de la Expresión Génica , Humanos , Leucocitos Mononucleares/inmunología , Leucocitos Mononucleares/microbiología , MicroARNs/genética
11.
Ying Yong Sheng Tai Xue Bao ; 34(10): 2871-2880, 2023 Oct.
Artículo en Zh | MEDLINE | ID: mdl-37897296

RESUMEN

Vertical greenery system (VGS) is a sustainable solution to promote building energy saving and emission reduction, mitigate the urban heat island effect, as well as a crucial component of urban ecological construction. We summarized four main mechanisms of the thermal effects of VGSs, including shading effect, evapotranspiration effect, thermal insulation effect, and wind control effect. We elucidated the effects of VGSs on building cooling and energy saving, and analyzed the cooling effects of VGSs on plant canopy and outdoor ambient air, as well as their influence on mitigating the urban heat island effect. Based on available research on the thermal effects of VGSs, we identified key directions for future research, aiming to expedite the development of green cities and achieve carbon neutrality.


Asunto(s)
Frío , Calor , Ciudades , Viento , Plantas
12.
Neural Regen Res ; 18(9): 1968-1975, 2023 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-36926721

RESUMEN

Patients with age-related hearing loss face hearing difficulties in daily life. The causes of age-related hearing loss are complex and include changes in peripheral hearing, central processing, and cognitive-related abilities. Furthermore, the factors by which aging relates to hearing loss via changes in auditory processing ability are still unclear. In this cross-sectional study, we evaluated 27 older adults (over 60 years old) with age-related hearing loss, 21 older adults (over 60 years old) with normal hearing, and 30 younger subjects (18-30 years old) with normal hearing. We used the outcome of the upper-threshold test, including the time-compressed threshold and the speech recognition threshold in noisy conditions, as a behavioral indicator of auditory processing ability. We also used electroencephalography to identify presbycusis-related abnormalities in the brain while the participants were in a spontaneous resting state. The time-compressed threshold and speech recognition threshold data indicated significant differences among the groups. In patients with age-related hearing loss, information masking (babble noise) had a greater effect than energy masking (speech-shaped noise) on processing difficulties. In terms of resting-state electroencephalography signals, we observed enhanced frontal lobe (Brodmann's area, BA11) activation in the older adults with normal hearing compared with the younger participants with normal hearing, and greater activation in the parietal (BA7) and occipital (BA19) lobes in the individuals with age-related hearing loss compared with the younger adults. Our functional connection analysis suggested that compared with younger people, the older adults with normal hearing exhibited enhanced connections among networks, including the default mode network, sensorimotor network, cingulo-opercular network, occipital network, and frontoparietal network. These results suggest that both normal aging and the development of age-related hearing loss have a negative effect on advanced auditory processing capabilities and that hearing loss accelerates the decline in speech comprehension, especially in speech competition situations. Older adults with normal hearing may have increased compensatory attentional resource recruitment represented by the top-down active listening mechanism, while those with age-related hearing loss exhibit decompensation of network connections involving multisensory integration.

13.
Nat Plants ; 9(8): 1221-1235, 2023 08.
Artículo en Inglés | MEDLINE | ID: mdl-37550371

RESUMEN

The origin of domesticated Asian rice (Oryza sativa L.) has been controversial for more than half a century. The debates have focused on two leading hypotheses: a single domestication event in China or multiple domestication events in geographically separate areas. These two hypotheses differ in their predicted history of genes/alleles selected during domestication. Here we amassed a dataset of 1,578 resequenced genomes, including an expanded sample of wild rice from throughout its geographic range. We identified 993 selected genes that generated phylogenetic trees on which japonica and indica formed a monophyletic group, suggesting that the domestication alleles of these genes originated only once in either japonica or indica. Importantly, the domestication alleles of most selected genes (~80%) stemmed from wild rice in China, but the domestication alleles of a substantial minority of selected genes (~20%) originated from wild rice in South and Southeast Asia, demonstrating separate domestication events of Asian rice.


Asunto(s)
Domesticación , Oryza , Filogenia , Oryza/genética , China , Alelos
14.
Sci China Life Sci ; 63(11): 1714-1724, 2020 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-32318909

RESUMEN

It is of critical importance for our understanding of speciation process to determine the forms of reproductive isolation and their relative importance in species divergence. Oryza nivara and O. rufipogon are direct ancestors of Asian cultivated rice and a progenitor-daughter species pair. Investigating the reproductive isolation between them provides insights into plant speciation and helps understanding of the rice domestication. Here, we quantitatively measured the major components of reproductive isolation between the two species based on common garden and crossing experiments for three pairs of sympatric populations in Nepal, Cambodia and Laos. We revealed significant differences in the flowering times between species pairs, with O. nivara flowering much earlier than O. rufipogon. A very weak reduction in seed set but no reduction in F1 viability and fertility were detected for the crosses between species relative to those within species. Moreover, we detected asymmetrical compatibility between species and found that emasculation significantly decreased pollination success in O. nivara but not in O. rufipogon. Our study demonstrates that the divergence between O. nivara and O. rufipogon is maintained almost entirely by the difference in flowering times and suggests that differential flowering times contribute to both habitat preferences and reproductive isolation between species.


Asunto(s)
Flores/genética , Oryza/genética , Aislamiento Reproductivo , Adaptación Fisiológica , Especiación Genética , Variación Genética , Polinización , Semillas/genética
15.
Mol Plant Microbe Interact ; 21(4): 411-23, 2008 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-18321187

RESUMEN

RsmA is an RNA-binding protein functioning as a global post-transcriptional regulator of various cellular processes in bacteria and has been demonstrated to be an important virulence regulator in many animal bacterial pathogens. However, its function in other phytopathogenic bacteria is unclear, except for the Erwinia carotovora RsmA, which acts as a negative virulence regulator. In this work, we investigated the function of the rsmA-like gene, named rsmA(Xcc), of the phytopathogen Xanthomonas campestris pv. campestris. Deletion of rsmA(Xcc) resulted in complete loss of virulence on the host plant Chinese radish, hypersensitive response on the nonhost plant pepper ECW-10R, and motility on the surface of an agar plate. The rsmA(Xcc) mutant displayed a significant reduction in the production of extracellular amylase, endoglucanase, and polysaccharide, but a significant increase in intracellular glycogen accumulation and an enhanced bacterial aggregation and cell adhesion. Microarray hybridization and semiquantitative reverse-transcription polymerase chain reaction analysis showed that deletion of rsmA(Xcc) led to significantly reduced expression of genes encoding the type III secretion system (T3SS), T3SS-effectors, and the bacterial aggregate dispersing enzyme endo-beta-1,4-mannanase. These results suggest that rsmA(Xcc) is involved in the control of various cellular processes, including pathogenesis of X. campestris pv. campestris.


Asunto(s)
Proteínas Bacterianas/genética , Regulación Bacteriana de la Expresión Génica , Mutación , Xanthomonas campestris/genética , Secuencia de Aminoácidos , Amilasas/metabolismo , Proteínas Bacterianas/fisiología , Celulasa/metabolismo , Glucógeno/metabolismo , Modelos Genéticos , Datos de Secuencia Molecular , Análisis de Secuencia por Matrices de Oligonucleótidos , Polisacáridos/metabolismo , Raphanus/crecimiento & desarrollo , Raphanus/microbiología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Homología de Secuencia de Aminoácido , Virulencia/genética , Xanthomonas campestris/metabolismo , Xanthomonas campestris/patogenicidad
16.
Ecol Evol ; 8(23): 12056-12065, 2018 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-30598799

RESUMEN

Genetic diversity and population genetic structure of the wild rice species Oryza rufipogon and O. nivara in Sri Lanka were studied using 33 microsatellite markers. A total of 315 individuals of 11 natural populations collected from the wet, intermediate, and dry zones of the country were used in the study. We found a moderate to high level of genetic diversity at the population level, with the polymorphic loci (P) ranging from 60.6% to 100% (average 81.8%) and the expected heterozygosity (H E) varying from 0.294 to 0.481 (average 0.369). A significant genetic differentiation between species and strong genetic structure within species were also observed. Based on species distribution modeling, we detected the dynamics of the preferred habitats for the two species in Sri Lanka and demonstrated that both O. rufipogon and O. nivara populations have expanded substantially since the last internal glacial. In addition, we showed that the geographical distribution of the two species corresponded to the climate zones and identified a few of key environmental variables that contribute to the distribution of the two species, implying the potential mechanism for ecological adaptation of these two species in Sri Lanka. These studies provided important insights into the population genetics and evolution of these wild species in Sri Lanka and are of great significance to the in situ conservation and utilization of these wild resources in genetic improvement of rice.

17.
PLoS One ; 10(7): e0134743, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26230627

RESUMEN

Real-time polymerase chain reaction (RT-PCR) enables effective and sensitive screening for infectious risk in the field of blood safety. However, when using RT-PCR to detect bacterial contamination, several intractable points must be considered, one of which is the lack of appropriate quality control. In this study, we developed a simplified RT-PCR assay in which the same primer set and two distinct probes were used to detect both, an internal reference control and the target in a reaction. The copy number of the internal reference control represents the positive detection limit of the assay; therefore, when the threshold-cycle value of the target is less than or equal to that of the internal reference control, the result obtained for the target can be considered to be a true positive. When human gDNA was spiked with Escherichia coli gDNA and the detection limit for the internal reference control was set to five copies, the measured detection limit for E. coli gDNA was two copies. The internal reference control duplex RT-PCR assay showed high efficiency (0.91-1.02), high linearity (R2 > 0.99), and good reproducibility in intra- and inter-assay comparisons. Lastly, when human platelet-rich plasma samples were spiked with E. coli or other bacterial species, all species were detected efficiently, and the results of a two-sample pooled t test showed that the limit of detection for E. coli was 1 cfu/mL. Here, we present a synthetic internal reference control molecule and a new statistical method for improving the reliability of RT-PCR assays when screening for bacterial contamination in blood products.


Asunto(s)
Bacteriemia/microbiología , Bacterias/aislamiento & purificación , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Bacterias/clasificación , Bacterias/genética , Secuencia de Bases , ADN Bacteriano/análisis , Humanos , Límite de Detección , Datos de Secuencia Molecular , Homología de Secuencia de Aminoácido
18.
FEMS Microbiol Lett ; 323(2): 180-7, 2011 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-22092718

RESUMEN

Mip (macrophage infectivity potentiator) and Mip-like proteins have been demonstrated to be involved in virulence of several animal pathogens, but as yet none of their native bacterial targets has been identified. Our previous work demonstrated that the Mip-like protein found in the plant pathogen Xanthomonas campestris pv. campestris (Xcc) (hereafter called Mip(Xcc)) is also involved in virulence. Inactivation of the mip(Xcc) gene leads to a significant reduction in exopolysaccharide production and extracellular protease activity via an unknown mechanism. The Xcc genome encodes six extracellular proteases, all of which are secreted via the type II secretion system. The serine protease PrtA makes the largest contribution to Xcc's total extracellular proteolytic activity. In this study, Western blotting analysis demonstrated that Mip(Xcc) was located in the periplasm. Bacterial two-hybrid and far-Western analysis indicated that Mip(Xcc) interacted with PrtA directly. Purified Mip(Xcc) was found to be able to rescue the protease activity of periplasmic proteins extracted from the mip(Xcc) mutant. These findings show that Mip(Xcc) plays a role in the maturation of PrtA, which is the novel native target for at least one Mip or Mip-like protein.


Asunto(s)
Proteínas Bacterianas/metabolismo , Péptido Hidrolasas/metabolismo , Factores de Virulencia/metabolismo , Xanthomonas campestris/enzimología , Proteínas Bacterianas/genética , Far-Western Blotting , Western Blotting , Técnicas de Inactivación de Genes , Péptido Hidrolasas/genética , Proteínas Periplasmáticas/genética , Proteínas Periplasmáticas/metabolismo , Unión Proteica , Mapeo de Interacción de Proteínas , Técnicas del Sistema de Dos Híbridos , Factores de Virulencia/genética , Xanthomonas campestris/genética
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