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Tree JJ Kidd SP Jennings MP McEwan AG 《Biochemical and biophysical research communications》2005,328(4):1205-1210
The cueO gene of Escherichia coli encodes a multi-copper oxidase, which contributes to copper tolerance in this bacterium. It was observed that a cueO mutant was highly sensitive to killing by copper ions when cells were grown on defined minimal media. Copper sensitivity was correlated with accumulation of copper in the mutant strain. Growth of the cueO mutant in the presence of copper could be restored by addition of divalent zinc and manganese ions or ferrous iron but not by other first row transition metal ions or magnesium ions. Copper toxicity towards a cueO mutant could also be suppressed by addition of the superoxide quencher 1,2-dihydroxybenzene-3,5-disulfonic acid (tiron), suggesting that a primary cause of copper toxicity is the copper-catalyzed production of superoxide anions in the cytoplasm. 相似文献
3.
Despite the power of sequencing and of emerging high-throughput technologies to collect data rapidly, the definitive functional characterization of unknown genes still requires biochemical and genetic analysis in case-by-case studies. This often involves the deletion of target genes and phenotypic characterization of the deletants. We describe here modifications of an existing deletion method which facilitates the deletion process and enables convenient analysis of the expression properties of the target gene by replacing it with an FRT-lacZ-aph-P(lac)-FRT cassette. The lacZ gene specifically reports the activity of the deleted gene and therefore allows the determination of the conditions under which it is actively expressed. The aph gene, encoding resistance to kanamycin, provides a selectable means of transducing a deleted locus between strains so that the deletion can be combined with other relevant mutations. The lac promoter helps to overcome possible polar effects on downstream genes within an operon. Because the cassette is flanked by two directly repeated FRT sites, the cassette can be excised by the Flp recombinase provided in trans. Removing the cassette leaves an in-frame deletion with a short scar which should not interfere with downstream expression. Replacements of yacF, yacG, yacH, yacK (cueO), yacL, ruvA, ruvB, yabB, and yabC made with the cassette were used to verify its properties. 相似文献
4.
Type 1 fimbriae have been implicated as virulence factors in animal models of urinary tract infection (UTI), but the function in human disease remains unclear. This study used a human challenge model to examine if type 1 fimbriae trigger inflammation in the urinary tract. The asymptomatic bacteriuria strain Escherichia coli 83972, which fails to express type 1 fimbriae, due to a 4.25 kb fimB-fimD deletion, was reconstituted with a functional fim gene cluster and fimbrial expression was monitored through a gfp reporter. Each patient was inoculated with the fim+ or fim- variants on separate occasions, and the host response to type 1 fimbriae was quantified by intraindividual comparisons of the responses to the fim+ or fim- isogens, using cytokines and neutrophils as end-points. Type 1 fimbriae did not promote inflammation and adherence was poor, as examined on exfoliated cells in urine. This was unexpected, as type 1 fimbriae enhanced the inflammatory response to the same strain in the murine urinary tract and as P fimbrial expression by E. coli 83972 enhances adherence and inflammation in challenged patients. We conclude that type 1 fimbriae do not contribute to the mucosal inflammatory response in the human urinary tract. 相似文献
5.
The antimicrobial peptide cathelicidin protects the urinary tract against invasive bacterial infection 总被引:1,自引:0,他引:1
Chromek M Slamová Z Bergman P Kovács L Podracká L Ehrén I Hökfelt T Gudmundsson GH Gallo RL Agerberth B Brauner A 《Nature medicine》2006,12(6):636-641
The urinary tract functions in close proximity to the outside environment, yet must remain free of microbial colonization to avoid disease. The mechanisms for establishing an antimicrobial barrier in this area are not completely understood. Here, we describe the production and function of the cathelicidin antimicrobial peptides LL-37, its precursor hCAP-18 and its ortholog CRAMP in epithelial cells of human and mouse urinary tract, respectively. Bacterial contact with epithelial cells resulted in rapid production and secretion of the respective peptides, and in humans LL-37/hCAP-18 was released into urine. Epithelium-derived cathelicidin substantially contributed to the protection of the urinary tract against infection, as shown using CRAMP-deficient and neutrophil-depleted mice. In addition, clinical E. coli strains that were more resistant to LL-37 caused more severe urinary tract infections than did susceptible strains. Thus, cathelicidin seems to be a key factor in mucosal immunity of the urinary tract. 相似文献
6.
Pseudomonas aeruginosa is an opportunistic human pathogen, which can cause severe urinary tract infections (UTIs). Because of the high intrinsic
antibiotic resistance of P. aeruginosa and its ability to develop new resistances during antibiotic treatment, these infections are difficult to eradicate. The
antibiotic susceptibility of 32 P. aeruginosa isolates from acute and chronic UTIs were analysed under standardized conditions showing 19% multi-drug resistant strains.
Furthermore, the antibiotic tolerance of two P. aeruginosa strains to ciprofloxacin and tobramycin was analysed under urinary tract-relevant conditions which considered nutrient composition,
biofilm growth, growth phase, and oxygen concentration. These conditions significantly enhance the antibiotic tolerance of
P. aeruginosa up to 6000-fold indicating an adaptation of the bacterium to the specific conditions present in the urinary tract. This reversible
phenomenon is possibly due to the increased formation of persister cells and is based on iron limitation in artificial urine.
The results suggest that the general high antibiotic resistance of P. aeruginosa urinary tract isolates together with the increasing tolerance of P. aeruginosa grown under urinary tract conditions decrease the efficiency of antibiotic treatment of UTIs. 相似文献
7.
Mutations in the receptor tyrosine kinase RET are associated with congenital anomalies of kidneys or urinary tract (CAKUT). RET tyrosine Y1015 is the docking site for PLCγ, a major regulator of RET signaling. Abrogating signaling via Y1015 causes CAKUT that are markedly different than renal agenesis in Ret-null or RetY1062F mutant mice. We performed analysis of Y1015F mutant upper and lower urinary tracts in mice to delineate its molecular and developmental roles during early urinary tract formation. We found that the degeneration of the common nephric ducts (CND), the caudal-most Wolffian duct (WD) segment, depends on Y1015 signals. The CNDs in Y1015F mutants persist owing to increased proliferation and reduced apoptosis, and showed abundance of phospho-ERK-positive cells. In the upper urinary tract, the Y1015 signals are required for proper patterning of the mesonephros and metanephros. Timely regression of mesonephric mesenchyme and proper demarcation of mesonephric and metanephric mesenchyme from the WD depends on RetY1015 signaling. We show that the mechanism of de novo ectopic budding is via increased ERK activity due to abnormal mesenchymal GDNF expression. Although reduction in GDNF dosage improved CAKUT it did not affect delayed mesenchyme regression. Experiments using whole-mount immunofluorescence confocal microscopy and explants cultures of early embryos with ERK-specific inhibitors suggest an imbalance between increased proliferation, decreased apoptosis and increased ERK activity as a mechanism for WD defects in RetY1015F mice. Our work demonstrates novel inhibitory roles of RetY1015 and provides a possible mechanistic explanation for some of the confounding broad range phenotypes in individuals with CAKUT. 相似文献
8.
Spencer JD Hains DS Porter E Bevins CL DiRosario J Becknell B Wang H Schwaderer AL 《PloS one》2012,7(2):e31712
Background
The mechanisms that maintain sterility in the urinary tract are incompletely understood. Recent studies have implicated the importance of antimicrobial peptides (AMP) in protecting the urinary tract from infection. Here, we characterize the expression and relevance of the AMP human alpha-defensin 5 (HD5) in the human kidney and urinary tract in normal and infected subjects.Methodology/Principal Findings
Using RNA isolated from human kidney, ureter, and bladder tissue, we performed quantitative real-time PCR to show that DEFA5, the gene encoding HD5, is constitutively expressed throughout the urinary tract. With pyelonephritis, DEFA5 expression significantly increased in the kidney. Using immunoblot analysis, HD5 production also increased with pyelonephritis. Immunostaining localized HD5 to the urothelium of the bladder and ureter. In the kidney, HD5 was primarily produced in the distal nephron and collecting tubules. Using immunoblot and ELISA assays, HD5 was not routinely detected in non-infected human urine samples while mean urinary HD5 production increased with E.coli urinary tract infection.Conclusions/Significance
DEFA5 is expressed throughout the urinary tract in non-infected subjects. Specifically, HD5 is expressed throughout the urothelium of the lower urinary tract and in the collecting tubules of the kidney. With infection, HD5 expression increases in the kidney and levels become detectable in the urine. To our knowledge, our findings represent the first to quantitate HD5 expression and production in the human kidney. Moreover, this is the first report to detect the presence of HD5 in infected urine samples. Our results suggest that HD5 may have an important role in maintaining urinary tract sterility. 相似文献9.
J. Steensberg E. D. Bartels H. Bay-Nielsen E. Fan?e T. Hede 《BMJ (Clinical research ed.)》1969,4(5680):390-394
During a one-year morbidity survey of urinary tract diseases in general practice 741 cases were diagnosed. Only about half of all the patients with symptoms of urinary tract infection had significant bacteriuria. In young women urinary tract infections and symptoms from the urinary tract without bacteriuria—in particular urethritis—were found to predominate. In middle-aged women, the urinary tract symptoms were ascribed increasingly to genital prolapse, while incidence of urolithiasis was the highest in any group, and urinary tract infections became less frequent. The prevalence of urinary tract infection showed another increase in elderly women, and recurrent/chronic pyelonephritis, which occurs with a steadily increasing prevalence throughout all age groups, became common.In younger male urological patients diseases with symptoms of urinary tract infection without bacteriuria were predominant, whereas prostatitis and urinary tract infections were less frequent. In middle-aged men, urolithiasis was especially frequent, while an increasing proportion of elderly men had prostatic hypertrophy, urinary tract infections, and recurrent/chronic pyelonephritis. 相似文献
10.
尿路上皮癌(urothelial carcinoma,uc)是泌尿系统最常见恶性肿瘤之一,早期诊断是提高该类疾病疗效的关键所在,荧光原位杂交(fluorescencein situ hybridization,FISH)通过尿液来检测UC,具有快速、无创伤性、敏感度高和特异性强等优点。FISH提高了尿细胞学在低级别或浅表性膀胱UC诊断的敏感性,且减少了血尿、尿路感染及膀胱内灌注治疗等对细胞形态的影响而引起的假阳性,提高检测的特异性。对于诊断上尿路UC,FISH的敏感性与特异性更高。膀胱UC患者9号染色体p16抑癌基因丢失与复发明显相关。FISH既能预测膀胱UC的复发性,更能监测UC的复发,但仍需大样本、多中心的前瞻性研究。本文将FISH在膀胱UC、上尿路UC早期诊断以及膀胱UC术后监测等方面的临床应用研究报道进行综述。 相似文献
11.
V M Korshunov F Z Nizamov N P Ivanova A V Ukhin Iu M Zakhmatov 《Zhurnal mikrobiologii, epidemiologii, i immunobiologii》1988,(4):8-13
The oral administration of microflora, isolated from the urinary tract or feces of urological patients, into the abacterial digestive tract of mice kept under the conditions of germ-free environment leads to the formation of the donors' microflora in the intestine of the recipient animals. Such recipient mice can be used as models for the rapid and highly reliable determination of the sensitivity of microflora in the patient's intestine and urinary tract to different antimicrobial preparations. In different patients such microflora, though similar in its qualitative and quantitative composition, differs in sensitivity to different antimicrobial preparations and their combinations. For the effective treatment of patients with chronic urinary tract infections a preliminary in vivo determination of the sensitivity of intestinal and urinary microflora to different combinations of antimicrobial preparations, carried out with the use of germ-free techniques, is necessary. 相似文献
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S. M. Hussain Qadri Fouad Al Dayel Michael J. Strampfer Burke A. Cunha 《Mycopathologia》1988,104(2):99-101
A case of urinary tract infection due to Hansenula anomala is reported. The infection occurred in a cadaver kidney transplant patient who was receiving immuno-suppression therapy. Survey of the literature revealed that human infections due to this organism are rare and its causal relationship in urinary tract infection has not been previously reported. 相似文献
14.
Over three years all infants in this hospital found to have an abnormality of the urinary tract on antenatal scanning were followed up after delivery with contrast radiography. Disease of the renal tract was confirmed in 17 of 20 infants. Of the 15 survivors, 12 underwent surgery in the first year of life. Abnormality of the fetal urinary tract detected by ultrasound scanning seems to be an important indicator of disease of the renal tract. Before its use is extended, however, further assessment of the benefit of antenatal diagnosis and of the best time to scan is required. 相似文献
15.
The diagnostic accuracy of sputum and urine cytology 总被引:1,自引:0,他引:1
W H Kern 《Acta cytologica》1988,32(5):651-654
The diagnostic accuracies of sputum and urinary cytology were examined in series spanning more than 20 years. The sensitivity and respiratory tract cytology in 1,289 patients with subsequently proven lung cancer was 69% while that or urine cytology in 860 patients with urinary tract cancer was 77%. The specificities were 96% for lung cancer and 97% for urinary tract cancer. Neither procedure was widely used for routine screening, but diagnostic cytology played an important part in providing a definite morphologic diagnosis in many of these cases. Urinary cytology was also very effective in the follow-up of patients with treated bladder cancer because of its high sensitivity for detecting carcinoma in situ. 相似文献
16.
Bubendorf L 《Acta cytologica》2011,55(2):113-119
Urinary cytology has a well-established role in the detection and monitoring of urothelial carcinoma. The main strength of cytology is the high specificity for high-grade urothelial carcinoma and carcinoma in situ, but it has a low sensitivity for low-grade, non-invasive tumors. There are several other limitations of cytology. Cytology of the upper urinary tract and after intravesical therapy with bacillus Calmette-Guerin is notoriously difficult to interpret. In addition, there is a poorly defined but commonly used category of atypical cytology of uncertain significance. The UroVysion multiprobe fluorescence in situ hybridization has emerged as a helpful tool to address these limitations. It consists of fluorescently labeled DNA probes to detect increased copy numbers (polysomy) of the chromosomes 3, 7 and 17 and deletion of 9p21, the site of the P16 tumor suppressor gene. Multiple studies have shown that fluorescence in situ hybridization in voided urine and washing specimens can help in patient management due to its superior sensitivity over cytology in different situations. It can be particularly useful to clarify equivocal cytological findings. However, some aspects remain to be further addressed including cost efficiency, optimal cut-off values and the true performance under real-life conditions. 相似文献
17.
Autosomal recessive Wolfram syndrome associated with an 8.5-kb mtDNA single deletion. 总被引:3,自引:0,他引:3
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A. Barrientos J. Casademont A. Saiz F. Cardellach V. Volpini A. Solans E. Tolosa A. Urbano-Marquez X. Estivill V. Nunes 《American journal of human genetics》1996,58(5):963-970
Wolfram syndrome (MIM 222300) is characterized by optic atrophy, diabetes mellitus, diabetes insipidus, neurosensory hearing loss, urinary tract abnormalities, and neurological dysfunction. The association of clinical manifestations in tissues and organs unrelated functionally or embryologically suggested the possibility of a mitochondrial implication in the disease, which has been demonstrated in two sporadic cases. Nonetheless, familial studies suggested an autosomal recessive mode of transmission, and recent data demonstrated linkage with markers on the short arm of human chromosome 4. The patient reported here, as well as her parents and unaffected sister, carried a heteroplasmic 8.5-kb deletion in mtDNA. The deletion accounted for 23% of mitochondrial genomes in lymphocytes from the patient and approximately 5% in the tissues studied from members of her family. The presence of the deletion in the patient in a proportion higher than in her unaffected parents suggests a putative defect in a nuclear gene that acts at the mitochondrial level. 相似文献
18.
Sarah B. Daly Emma Hilton Richard A. Kammerer Bronwyn Kerr Dian Donnai Berk Burgu Murat Derbent Willie Reardon Stavit Shalev Adrian S. Woolf William G. Newman 《American journal of human genetics》2010,86(6):963-674
Urinary voiding dysfunction in childhood, manifesting as incontinence, dysuria, and urinary frequency, is a common condition. Urofacial syndrome (UFS) is a rare autosomal recessive disease characterized by facial grimacing when attempting to smile and failure of the urinary bladder to void completely despite a lack of anatomical bladder outflow obstruction or overt neurological damage. UFS individuals often have reflux of infected urine from the bladder to the upper renal tract, with a risk of kidney damage and renal failure. Whole-genome SNP mapping in one affected individual defined an autozygous region of 16 Mb on chromosome 10q23-q24, within which a 10 kb deletion encompassing exons 8 and 9 of HPSE2 was identified. Homozygous exonic deletions, nonsense mutations, and frameshift mutations in five further unrelated families confirmed HPSE2 as the causative gene for UFS. Mutations were not identified in four additional UFS patients, indicating genetic heterogeneity. We show that HPSE2 is expressed in the fetal and adult central nervous system, where it might be implicated in controlling facial expression and urinary voiding, and also in bladder smooth muscle, consistent with a role in renal tract morphology and function. Our findings have broader implications for understanding the genetic basis of lower renal tract malformations and voiding dysfunction. 相似文献
19.
D F Kinane C C Blackwell R P Brettle D M Weir F P Winstanley R A Elton 《BMJ (Clinical research ed.)》1982,285(6334):7-9
ABO blood group and secretor state was determined in 319 women with recurrent urinary tract infection and compared with those of a control group of 334 women of similar age ranges. Women of blood groups B and AB who are non-secretors of blood group substances showed a significant relative risk of recurrent urinary tract infection of 3.12 (95% confidence limits, 1.49 and 6.52) in comparison with other types. This appears to be a genuine example of synergy in which absence of anti-B isohaemagglutinin and secretor substances combines to give an increased risk of recurrent urinary tract infection. Determination of blood group and secretor state may provide additional information in identifying those at risk. 相似文献
20.
K S Joseph K N Brahmadathan Sulochana Abraham Abraham Joseph 《BMJ (Clinical research ed.)》1988,296(6626):906-907
Urinary tract infection in pregnancy has not been adequately dealt with in developing countries, though its consequences are well recognised. This is primarily because of constraints on resources coupled with a lack of technological infrastructure. An evaluation of the Griess test for the mass screening of urinary tract infection among antenatal women was carried out prospectively using a case-control method.The Griess test was found to be a valid, reliable, and economical screening test for urinary tract infection which can be integrated into a primary maternal and child health care programme. 相似文献