Discussions over the past three decades, scientists have identified huge effort to create animal models of NEC. Persecution of disease states characterized by necrosis of the intestine, increased inflammatory mediators, and the open destruction of histological bore extensive knowledge base about the characteristics of the final stages of NEC. However, these models have some limited understanding of the early images required for the progression of NEC from relatively mild intestinal inflammation in a state of true necrosis. To expand our knowledge of early triggers NEC, we describe our efforts to investigate the pathogenesis of NEC in the early stages of the disease. Despite the diagnosis of NEC based on well-confirmed histological point system, with no gross necrosis was found in one of our animals, and animals do not have ratings higher than the NEC 2. This suggests that the disease is in a relatively early stage, preceding the development of gross necrosis and histological characteristics of the destruction of more advanced states of disease (stage 4). Our further analysis of the factors commonly implicated in the pathogenesis of NEC (bacteria and inflammatory mediators), and our SPS shows that even in the earliest stages of the disease, a unique intestinal bacterial community structure exists in animals with NEC compared with the disease without animals. Given that mice in our model was the height of the inflammatory mediator typical late stage NPC, we can extrapolate that the increase in inflammatory mediators is later marker of advanced stages of disease. Taken together, these findings suggest that temporal relationships may exist between the intestinal microflora and inflammatory mediators, such that changes in gut bacteria may be associated with early NEC pathogenesis in inflammatory mediator height can occur later in the progression of the disease. In addition to the conclusion that changes in gut microflora associated with early stages of NEC, our work uses non-culture technology for detection of specific pathogens associated with disease. Recent studies show that only about 5% of all known species of bacteria and 30BЂ "40% of all bacteria associated with human body successfully cultivated with traditional culture-based laboratory methods. Our use of new molecular approaches to bacterial identification, such as 454 16S ribosomal pirosekvenirovaniya of subunits, very conservative area is now all bacterial cells with Hipervariatyvna region, which differs in different species of bacteria, thus allowing in-depth analysis of molecular organisms present in the disease without reliance on traditional culture-based methods. Interestingly, our work demonstrates the increasing relative content of gram-negative microorganisms (
Citrobacter, Klebsiella
and
Tatumella) in a thick flora of animals with diseases compared to animals without disease. While the finding of increased gram-negative bacteria with NEC previously described in studies using traditional culture-based technologies, molecular mechanisms by which this class of bacteria may be uniquely well suited to play an important role in the pathophysiology of NEC remains unknown. Further investigation into the properties of gram-negative bacteria, which can provide NEC is important for understanding the pathophysiology of NEC. However, the mere presence of bacteria in disease offers a limited knowledge of changes in the genome of the bacteria that provide increased virulence and subsequent disease progression. Further experiments that use advanced molecular techniques to study metatranscriptome (whole community bacterial gene expression) bacteria associated with the early pathophysiology of NEC will provide new understanding of bacterial gene called to initiate disease. In addition, it is possible that changes in the host epithelial mucosa secondary to hypoxia, feeding and other factors to manipulate to create NEC can contribute to conditions in the lumen of the intestine, which promotes the survival advantage for certain types of bacteria. The measurement of complex host-microbe relationship of molecular genetic level will offer key understanding of the pathogenesis of NEC. Further refinement of our understanding of the pathophysiology of NEC early factors, along with increased availability of clinical 16S rRNA sequences will increase our ability to offer early therapeutic intervention for children considered at risk for NEC. Previous applications for 16S rRNA sequence of methods used in our experimental model is conducted in an effort to enrich our understanding of microbial ecology of premature babies. For example, in a study 23 infants who were born in 23BЂ "32 weeks of pregnancy, Mshvildadze et al found that while the overall profile of microbial origin in children with NEC did not differ from control children with NEC increased level
Citrobacter and Enterococcus compared with infants without NEC. Intestinal microflora structure differential between children and without NEC also was discovered that de la Cochetiere prospectively analyzed samples of stool from 3 children who developed NEC and 9, which does not define what
Clostridium perfringens are present in children who developed NEC but not able to control children. On the other hand, Miller and others have used 16S rRNA sequence based analysis of fecal microflora of children and found no differences between babies and without NEC. According to a young El Al in his review of potential biomarkers for NEC, continued use of 16S rRNA based sequencing offer new possibilities for the separation of the gut microbial community structures that contribute to the pathogenesis of NEC. Better understanding of these bacterial communities provide a novel understanding of how physicians may be able to manage gut microflora with antibiotics or probiotics target for the development of preventive therapy for NEC. Whereas the realistic problems of high multidrug-resistant bacteria secondary to aggressive use of antibiotics, such focused approaches to treatment may be better than the current practice of empirical administration of broad-spectrum antibiotics for suspected NEC. In addition, the transition to prevention rather than supportive therapy in NEC will allow physicians to more accurately and aggressively treat early NEC, that the progression of disease in developed countries the disease requiring surgical intervention or drainage could have been avoided. While we hope that the results of this work to deepen understanding of the microbial contribution to the pathogenesis of NEC and suggest new approaches to targeted therapies, this study is not without limitations. Although we conclude that the gram-negative bacteria have an increased abundance of animals with NEC based on the literature, relatively small sample size in our study, requires further study to ensure consistent reproducibility. In particular, our DNA / RNA analysis was limited in that instead of analyzing the full group of mice, we estimated eight animals. We elected to include only those animals with the highest quality RNA to provide the most accurate results, however, this study will be strengthened by increasing the sample size. Given the numerous experimental models of NEC, our results could be enhanced if they can be reproduced in other models. To replicate our consistent findings emerging direct clinical application is limited. In addition, our studies of inflammatory mediator expression may be enhanced by using techniques such as laser capture microscopy to evaluate expression of mRNA levels in epithelial cells, but not generally at the level of tissues. Finally, our work uses a 7-day animals, but not newborns or premature animal that may be more concerned with the development of similar children suffering from NEC. The older animals were used in a manner consistent with generation beginning of the disease in premature infants or animals is difficult because the influence of experimental conditions, which generates NEC is a lot of young animals to progress quickly to more complex diseases. However, capturing the early progression of NEC in premature infants and animals will allow us to more clearly determine the influence of microbes on immature intestinal physiology. Further experiments will be used for strattera 40mg this purpose. In addition, the use of older animals may have accounted for our slightly lower NEC. Several researchers describe the incidence of NEC, which ranges from 40BЂ "60%, but these studies used newborn animals. Although the meet NEC was established in our animals, 7 days old animals may be more resistant to damage caused by this protocol. This led to the reduction of vulnerability may be made for our slightly lower NEC. In this manuscript, we believe that research to the NEC gross necrosis and almost deadly disease state allows novel understanding of the mediators of early disease progression. Our results show that, despite the absence of inflammatory mediator high in mice with early NEC, intestinal microbial community structure did not differ in animals with early stage disease compared to animals without disease. This can be illustrated by the temporary relationship between bacteria and inflammatory mediators that altered intestinal microflora is associated with early pathogenesis NEC while inflammatory mediators that later downstream marker of disease progression. Further studies to examine the complex, bidirectional communication between host epithelial cells, intestinal microbes, and immature gut physiology in early NEC will offer unique insight into the etiology of NEC and potential new therapeutic targets to prevent progression of lung inflammation severe necrosis of the intestine . .
