Phylogenic classification and virulence genes profiles of uropathogenic E. coli and diarrhegenic E. coli strains isolated from community acquired infections.

The emergence of E.coli strains displaying patterns of virulence genes from different pathotypes shows that the current classification of E.coli pathotypes may be not enough, the study aimed to compare the phylogenetic groups and urovirulence genes of uropathogenic Escherichia coli (UPEC) and diarrheagenic E.coli (DEC) strains to extend the knowledge of E.coli classification into different pathotypes. A total of 173 UPEC and DEC strains were examined for phylogenetic typing and urovirulence genes by PCR amplifications. In contrast to most reports, phylogenetic group A was the most prevalent in both UPEC and DEC strains, followed by B2 group. Amplification assays revealed that 89.32% and 94.29% of UPEC and DEC strains, respectively, carried at least one of the urovirulence genes, 49.5% and 31.4% of UPEC and DEC strains, respectively, carried ≥ 2 of the urovirulence genes, fim H gene was the most prevalent (66.9% and 91.4%) in UPEC and DEC strains respectively. Twenty different patterns of virulence genes were identified in UPEC while 5 different patterns in DEC strains. Strains with combined virulence patterns of four or five genes were belonged to phylogenetic group B2. Our finding showed a closer relationship between the DEC and UPEC, so raised the suggestion that some DEC strains might be potential uropathogens. These findings also provide different insights into the phylogenetic classification of E. coli as pathogenic or commensals where group A can be an important pathogenic type as well as into the classification as intestinal or extra- intestinal virulence factors

Modified Adenovirus Reduces De Novo Peritoneal Adhesions in Rats and Limits Off-Target Transfection. Role of EZH2 in Adhesion Formation

Aim of the study: Adenovector encoding tissue plasminogen activator (tPA) was shown to reduce experimental peritoneal adhesion. We investigated the targeting potential of our modified adenovector, its ability to reduce adhesions and the epigenetic role of histone methyltransferase EZH2 in adhesion formation. Materials and methods: Control lacZ, nonmodified tPA or modified tPA vectors were instilled in the peritoneal cavity after injury in de novo adhesions or after lysis of adhesions in recurrent adhesions. Adhesion severity was scored and adhesions and liver tissues were examined for adenovirus E4 gene and tPA mRNA expression. Levels of tPA, plasminogen activator inhibitor-1 (PAI-1), transforming growth factor-β1 (TGF-β1), and EZH2 expression were measured. Results: E4 transcripts were detected in adhesions of nonmodified and modified and in livers of nonmodified but not in livers of modified de novo adhesions. Both nonmodified (p = 0.021) and modified vectors (p = 0.036) reduced the severity of de novo adhesions compared to lacZ vector. Levels of tPA in nonmodified (p = 0.021) and modified adhesions (p = 0.001) were elevated while PAI-1 (p = 0.013 and p = 0.001, respectively) and TGF-β1 levels (p = 0.002 and p = 0.016, respectively) were reduced compared with lacZ group. All vectors were not expressed in recurrent adhesions and severity score were not different among groups. EZH2 levels were elevated in de novo nontreated (p = 0.001) and was further increased in recurrent (p = 0.001) nontreated adhesions compared with noninjured peritoneum. Conclusion: Modified adenovirus successfully targeted de novo adhesions but not liver tissues and reduced the severity of de novo adhesions. EZH2 is involved in the development and progression of peritoneal adhesions

3D CT volumetric analysis of the lung in COPD patients: Comparison with pulmonary function tests

Background: Retro and prospective comparative analytical study will be done over a 2-year period on patients with a multidisciplinary diagnosis of chronic obstructive pulmonary disease at the radiology department of Ain Shams University Hospitals. The aim of the study: To assess the role of the new CT lung analysis in quantitative assessment of chronic obstructive lung disease. The procedures: Patients will be subjected to CT chest without contrast then 3D models will be reconstructed using a software that allows automatic segmentation of each lung lobes and volume calculation, and giving each lung zone a Goddard score to determine the severity. The instruments: Patients will be assessed by the pulmonary function tests and body plethysmography. The results: The results will be collected and compared together. Data analysis will be performed using IBM statistical program for social science, version 22.0 (SPSS, 2013; IBM Corp., USA). Quantitative and qualitative data will be expressed as mean±SD, frequencies, and percentages. In addition, the relationship between total lung capacity (TLC), CT lung volume, and percentage LAA will be assessed and considered significant when the P-value is less than 0.05.&nbsp

Mutant MMP-9 and HGF Gene Transfer Enhance Resolution of CCl₄-Induced Liver Fibrosis in Rats: Role of ASH1 and EZH2 Methyltransferases Repression

<div><p>Hepatocyte growth factor (HGF) gene transfer inhibits liver fibrosis by regulating aberrant cellular functions, while mutant matrix metalloproteinase-9 (mMMP-9) enhances matrix degradation by neutralizing the elevated tissue inhibitor of metalloproteinase-1 (TIMP-1). It was shown that ASH1 and EZH2 methyltransferases are involved in development of liver fibrosis; however, their role in the resolution phase of liver fibrosis has not been investigated. This study evaluated the role of ASH1 and EZH2 in two mechanistically different therapeutic modalities, HGF and mMMP-9 gene transfer in CCl₄ induced rat liver fibrosis. Liver fibrosis was induced in rats with twice a week intraperitoneal injection of CCl₄ for 8 weeks. Adenovirus vectors encoding mMMP-9 or HGF genes were injected through tail vein at weeks six and seven and were sacrificed one week after the second injection. A healthy animal group was likewise injected with saline to serve as a negative control. Rats treated with mMMP-9 showed significantly lower fibrosis score, less Sirius red stained collagen area, reduced hydroxyproline and ALT concentration, decreased transforming growth factor beta 1 (TGF-β1) mRNA and lower labeling indices of α smooth muscle actin (α-SMA) and proliferating cell nuclear antigen (PCNA) stained cells compared with HGF- or saline-treated rats. Furthermore, TIMP-1 protein expression in mMMP-9 group was markedly reduced compared with all fibrotic groups. ASH1 and EZH2 protein expression was significantly elevated in fibrotic liver and significantly decreased in mMMP-9- and HGF-treated compared to saline-treated fibrotic livers with further reduction in the mMMP-9 group. Conclusion: Gene transfer of mMMP-9 and HGF reduced liver fibrosis in rats. ASH1 and EZH2 methyltransferases are significantly reduced in mMMP-9 and HGF treated rats which underlines the central role of these enzymes during fibrogenesis. Future studies should evaluate the role of selective pharmacologic inhibitors of ASH1 and EZH2 in resolution of liver fibrosis.</p></div

Effect of gene therapy on molecular markers of liver fibrosis.

<p>(A, B) Col1α1 mRNA expression was significantly elevated (*, p<0.001) in saline-treated livers compared with healthy livers (n = 10), and was significantly reduced with HGF (n = 14, #, p<0.05), mMMP-9 (n = 10, *, p<0.001) and HGF/mMMP-9 (n = 8, p<0.001) treatments compared with saline-treated fibrotic livers (n = 13). (C, D) α-SMA mRNA expression was significantly reduced with HGF (n = 14, p<0.001), mMMP-9 (n = 10, p<0.001) and HGF/mMMP-9 (n = 9, p<0.001) treatments compared with saline-treated fibrotic livers and was significantly elevated (p<0.001) in saline-treated fibrotic livers compared with healthy livers (n = 10). (E, F) TGF-β1 mRNA expression was significantly reduced with mMMP-9 (n = 10, p<0.01) and HGF/mMMP-9 (n = 9, p<0.01) treatment but not with HGF (n = 13, p = 0.87) compared with saline-treated fibrotic livers (n = 13). (G) TIMP-1 protein concentration was elevated (p<0.01) in saline-treated fibrotic livers (n = 9) and was further elevated in HGF-treated livers (n = 13) compared with healthy liver (n = 9). TIMP-1 concentration in fibrotic livers was significantly (p<0.001) decreased with mMMP-9 (n = 10) but not with HGF (p = 0.09) or with HGF/mMMP-9 (n = 9, p = 0.69) treatment. (H) Hydroxyproline concentration in saline-treated fibrotic livers (n = 10) was significantly elevated (p<0.001) compared with healthy livers (n = 10) and was significantly (p<0.001) decreased only with mMMP-9 treatment (n = 10) but not with HGF (n = 11, p = 0.86) or with HGF/mMMP-9 (n = 7, p = 0.98) treatments. (I) ALT serum concentration in saline-treated fibrotic rats (n = 14) was significantly elevated (p<0.001) compared with healthy rats (n = 10) and was significantly decreased with mMMP-9 (n = 10, p<0.001) and combined HGF/mMMP-9 (n = 9, p<0.01) but not with HGF (n = 14, p = 0.15) treatment.</p

Histopathological findings in rat liver sections stained with H&E and Sirius-red.

<p>Fibrosis score was significantly elevated (*, p<0.001) in saline-treated fibrotic (n = 14) compared with healthy livers (n = 10). Fibrosis scores of HGF-treated livers (n = 14) were not statistical different from those of saline-treated livers (p = 0.71). However, fibrosis score was significantly reduced (**, p<0.01) in mMMP-9-treated (n = 9) and (#, p<0.05) in HGF/mMMP-9-treated (n = 9) compared to saline-treated fibrotic livers. (Original magnification ×200). Sirius red stained collagen area percent was significantly elevated (p<0.001) in saline-treated fibrotic livers (n = 14) compared with healthy livers (n = 10). Collagen area percent was significantly reduced in mMMP-9-treated (n = 9, p<0.001)) and in HGF/mMMP-9-treated (n = 9, p<0.01) but not in HGF-treated (n = 13, p = 0.19) compared to saline-treated fibrotic livers. (Original magnification, ×100).</p

Analysis of ASH1 and EZH2 methyltransferases protein expression with Slot Blot.

<p>ASH1 (A, B) and EZH2 (C, D) protein expression were undetectable in the healthy livers. Both ASH1 and EZH2 protein expression markedly increased (p<0.01) following CCl₄ treatment for eight weeks. Treatment with HGF, mMMP-9 or combined HGF/mMMP-9 resulted in significant (p<0.01) reduction of both ASH1 and EZH2 protein expression in fibrotic livers. There was no significant difference in the ASH1 or EZH2 protein expression between the HGF-, mMMP-9-, or HGF/mMMP-9-treated fibrotic livers. (n = 6 per group).</p

Immunohistochemical staining for α-SMA and PCNA in the liver.

<p>Bar graphs represent α-SMA and PCNA labeling indices (percent of positively stained brown cytoplasm or brown nuclei, respectively in 10 successive fields). Both α-SMA and PCNA indices were significantly elevated (*, p<0.001) in saline-treated fibrotic (n = 14 both) compared with healthy livers (n = 10 both) and both were significantly reduced (*, p<0.001) in mMMP-9-treated (n = 10 both) and in HGF/mMMP-9-treated (n = 9 both, *, p<0.001 and #, p<0.01, respectively) versus saline-treated fibrotic livers. Both α-SMA and PCNA labeling indices were not significantly different in HGF-treated fibrotic (n = 14, p = 0.99 and p = 0.06, respectively) from that in saline-treated fibrotic livers. (Original magnification ×400).</p

Analysis of HGF and MMP-9 gene expression.

<p>(A, B) RT-PCR gel of HGF mRNA expression. HGF mRNA expression was significantly (*, p<0.001) elevated in the saline-treated fibrotic (n = 14) compared to saline-treated healthy livers (n = 10) and further elevated (#, p<0.01) in the HGF-treated fibrotic (n = 13) compared with the saline-treated fibrotic and mMM-9-treated fibrotic (n = 10) livers (*, p<0.01). (C) Analysis of HGF protein concentration in the liver tissues. HGF protein concentration was significantly (*, p<0.001) elevated in the liver of the saline-treated fibrotic livers (n = 11) compared to saline-treated healthy livers (n = 10) and more elevated in the HGF-treated fibrotic (n = 14) compared to the saline-treated fibrotic (#, p<0.05) and mMMP-9-treated fibrotic (n = 10) livers (**, p<0.01). (D) Analysis of MMP-9 protein concentration in the liver tissues. MMP-9 protein concentration was significantly elevated in the mMMP-9-treated (n = 10) compared with healthy (n = 9), saline-treated fibrotic (n = 13) livers, HGF-treated (n = 12) livers (*, p<0.01) and HGF/mMMP-9-treated (n = 6) livers (#, p<0.05).</p