The leukemia associated ETO nuclear repressor gene is regulated by the GATA-1 transcription factor in erythroid/megakaryocytic cells

BACKGROUND: The Eight-Twenty-One (ETO) nuclear co-repressor gene belongs to the ETO homologue family also containing Myeloid Translocation Gene on chromosome 16 (MTG16) and myeloid translocation Gene-Related protein 1 (MTGR1). By chromosomal translocations ETO and MTG16 become parts of fusion proteins characteristic of morphological variants of acute myeloid leukemia. Normal functions of ETO homologues have as yet not been examined. The goal of this work was to identify structural and functional promoter elements upstream of the coding sequence of the ETO gene in order to explore lineage-specific hematopoietic expression and get hints to function.RESULTS: A putative proximal ETO promoter was identified within 411 bp upstream of the transcription start site. Strong ETO promoter activity was specifically observed upon transfection of a promoter reporter construct into erythroid/megakaryocytic cells, which have endogeneous ETO gene activity. An evolutionary conserved region of 228 bp revealed potential cis-elements involved in transcription of ETO. Disruption of the evolutionary conserved GATA -636 consensus binding site repressed transactivation and disruption of the ETS1 -705 consensus binding site enhanced activity of the ETO promoter. The promoter was stimulated by overexpression of GATA-1 into erythroid/megakaryocytic cells. Electrophoretic mobility shift assay with erythroid/megakaryocytic cells showed specific binding of GATA-1 to the GATA -636 site. Furthermore, results from chromatin immunoprecipitation showed GATA-1 binding in vivo to the conserved region of the ETO promoter containing the -636 site. The results suggest that the GATA -636 site may have a role in activation of the ETO gene activity in cells with erythroid/megakaryocytic potential. Leukemia associated AML1-ETO strongly suppressed an ETO promoter reporter in erythroid/megakaryocytic cells.CONCLUSIONS: We demonstrate that the GATA-1 transcription factor binds and transactivates the ETO proximal promoter in an erythroid/megakaryocytic-specific manner. Thus, trans-acting factors that are essential in erythroid/megakaryocytic differentiation govern ETO expression

The renal pentad

Diabetes management is a comprehensive exercise which encompasses not only glycemic control, but vascular risk reduction as well. Accepted clinical models such as the glycemic pentad and metabolic pentad list the glucose related and metabolic aspects which influence ling term vascular outcomes. This paper describes a 'renal pentad' which consists of 5×2 easily measurable parameters, which influence renal outcomes. Renal function ,acute health concerns, chronic health concerns, glycemic control and comorbid concerns from the five components of this pentad. The 5 pointed rubric serves as a teaching and clinical tool, and assists in appropriate choice and targets of therapy in diabetic kidney disease

Kinetics of cytokine profile in response to Mycobacterium bovis BCG and Streptococcus pyogenes activated cells

The infection of epithelial cells is a necessary step for Mycobacterium bovis BCG dissemination, but the mechanism of mycobacterial epithelial interactions is not completely understood. Similarly, Streptococcus pyogenes is a strictly human pathogen that favorably colonizes the skin and the pharynx. Effective cytokine secretion is essential in order to fabricate a suitable inflammatory response against an infection. In this data article, the cytokine profile in BCG and S. pyogenes activated THP-1 cell line in media after the acute phase of infection by ELISA is described. The interleukin-8 level was increased in response to both BCG and S. pyogenes, but was quite prominent after 24 h and further increased upto 72 h post infection. On the other hand, an increase in IL-6 response to S. pyogenes was observed while there was no response to BCG even after 48 h of infection. A low level of TNF-α was detected upon BCG and S. pyogenes infection

The leukemia associated nuclear corepressor ETO homologue genes <it>MTG16 </it>and <it>MTGR1 </it>are regulated differently in hematopoietic cells

<p>Abstract</p> <p>Background</p> <p>MTG16, MTGR1 and ETO are nuclear transcriptional corepressors of the human ETO protein family. MTG16 is implicated in hematopoietic development and in controlling erythropoiesis/megakaryopoiesis. Furthermore, ETO homologue genes are 3'participants in leukemia fusions generated by chromosomal translocations responsible of hematopoietic dysregulation. We tried to identify structural and functional promoter elements of <it>MTG16 </it>and <it>MTGR1 </it>genes in order to find associations between their regulation and hematopoiesis.</p> <p>Results</p> <p>5' deletion examinations and luciferase reporter gene studies indicated that a 492 bp sequence upstream of the transcription start site is essential for transcriptional activity by the <it>MTG16 </it>promoter. The TATA- and CCAAT-less promoter with a GC box close to the start site showed strong reporter activity when examined in erythroid/megakaryocytic cells. Mutation of an evolutionary conserved GATA -301 consensus binding site repressed promoter function. Furthermore, results from <it>in vitro </it>antibody-enhanced electrophoretic mobility shift assay and <it>in vivo </it>chromatin immunoprecipitation indicated binding of GATA-1 to the GATA -301 site. A role of GATA-1 was also supported by transfection of small interfering RNA, which diminished <it>MTG16 </it>expression. Furthermore, expression of the transcription factor HERP2, which represses GATA-1, produced strong inhibition of the <it>MTG16 </it>promoter reporter consistent with a role of GATA-1 in transcriptional activation. The TATA-less and CCAAT-less <it>MTGR1 </it>promoter retained most of the transcriptional activity within a -308 to -207 bp region with a GC-box-rich sequence containing multiple SP1 binding sites reminiscent of a housekeeping gene with constitutive expression. However, mutations of individual SP1 binding sites did not repress promoter function; multiple active SP1 binding sites may be required to safeguard constitutive <it>MTGR1 </it>transcriptional activity. The observed repression of <it>MTG16</it>/<it>MTGR1 </it>promoters by the leukemia associated <it>AML1</it>-<it>ETO </it>fusion gene may have a role in hematopoietic dysfunction of leukemia.</p> <p>Conclusions</p> <p>An evolutionary conserved GATA binding site is critical in transcriptional regulation of the <it>MTG16 </it>promoter. In contrast, the <it>MTGR1 </it>gene depends on a GC-box-rich sequence for transcriptional regulation and possible ubiquitous expression. Our results demonstrate that the <it>ETO </it>homologue promoters are regulated differently consistent with hematopoietic cell-type- specific expression and function.</p

Inflammasomes and Their Role in Innate Immunity of Sexually Transmitted Infections

Inflammasomes are multiprotein complexes present in the cytosol as Pattern Recognition Receptors (PRRs) or as sensors of Damage Associated Molecular Patterns (DAMPs). After recognition of Microbe Associated Molecular Patterns (MAMPs) or host derived danger signals, Nucleotide oligomerization domain (NOD) like receptors oligomerize to form inflammasomes. The activation of inflammasomes results in an alarm, which is raised to alert adjacent cells through the processing and release of a number of other substrates present in the cytosol. A wide array of inflammasomes and their adapter molecules have been identified in the host’s innate immune system in response to a variety of pathogens. Components of specific pathogens activate different inflammasomes, which once activated in response to pathogen-induced infection, induce the activation of caspases and the release of mature forms of interleukin-1β (IL-1β) and IL-18. Identifying the mechanisms underlying pathogen-induced inflammasome activation is important if we are to develop novel therapeutic strategies to target STI related pathogens. This information is currently lacking in literature. In this review, we have discussed the role of various inflammasomes in sensing different sexually transmitted infections, as well as the beneficial or detrimental effects of inflammasome signaling in host resistance. Additionally we have discussed both canonical and noncanonical processing of IL-1β induced with respect to particular infections. Overall, these findings transform our understanding of both the basic biology and clinical relevance of inflammasomes

Data showing levels of interleukin-1β and nitric oxide in the plasma of uropathogenic E. coli infected UTI patients

Urinary tract infections (UTI) are a major cause of morbidity, affecting at least four million women worldwide, 65–75% of these infections are caused by Uropathogenic Escherichia coli (UPEC) (Foxman, 2010) [1]. Repertoire of virulence factors carried by UPEC provides the ability to precede urinary tract and additionally they provoke pro-inflammatory responses (Cirl et al., 2008; Verma et al., 2016) [2,3]. In context to UPEC infected UTI patients, the levels of pro-inflammatory cytokine IL-1β and enzymatic antioxidant nitric oxide (NO) have not been reported worldwide till date, including India. In this data article, we report for the first time the levels of IL-1β and nitric oxide in the plasma of UPEC infected UTI patients. Data includes a profile of pro-inflammatory cytokine IL-1β and NO in the plasma of the confirmed UPEC infected UTI patients (N = 30) versus healthy controls (N = 40) from the present pilot study. The levels of IL-1β in plasma were significantly higher (p < 0.0001) in patients (252.3 ± 6.49 pg/ml) as compared to healthy controls (127.6 ± 3.98 pg/ml), whereas plasma levels of NO were significantly lower (p < 0.0001) in UPEC infected UTI patients (60.29 ± 1.1 μM) as compared to healthy controls (106.3 ± 8.75 μM). Keywords: Uropathogenic Escherichia coli, Interleukin-1β, Nitric oxid

Alarming levels of antimicrobial resistance among sepsis patients admitted to ICU in a tertiary care hospital in India - a case control retrospective study

Abstract Background Hospital acquired infections (HAI) are principal threats to the patients of intensive care units. An increase in the antimicrobial resistance (AMR) observed in gram negative bacteria is a great challenge to deal with. HAI and AMR lead to prolonged hospitalization and additional doses of anti-microbial treatment affecting patient’s fitness and finances. Present study was undertaken to determine the pathotypes, genetic diversity and the antimicrobial resistance of E.coli in isolates from the patients admitted to intensive care unit at a tertiary care hospital in Delhi, India. Methods E.coli isolates (N = 77) obtained from the blood culture of patients diagnosed with sepsis and the isolates (N = 71) from the stool culture of patients admitted in intensive care unit (ICU) but not diagnosed with sepsis were investigated for their pathotypes, adherence patterns and genetic diversity by Enterobacterial Repeated Intergenic Consensus-polymerase chain reaction (ERIC-PCR). A Kirby-Bauer Disc diffusion test and antimicrobial susceptibility assays were performed according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. Extended-spectrum β-lactamase (ESBL) genes and sequence type 131 (ST131) clone were characterised genotypically by gene-specific PCRs. Results Pathotypes analysis revealed 46 and 16% of the blood E.coli isolates were ETEC and EAEC respectively, in contrast to the fecal isolates wherein 22% of the isolates were ETEC and 28.5% were EAEC. EPEC, STEC and EIEC pathotypes were not detected in blood or fecal isolates. Of all the isolates studied, more than 90% of the blood and 70% of the fecal isolates were found to be resistant to cephalosporins. On the other hand, 68% of blood and 44% of the fecal isolates were found to be ESBL producers. Interestingly 83% of the blood isolates contained CTX-M15, whereas only 21% of them contained CTX-M9 genes. On the other hand CTX-M15 genes were found in 90% and CTX-M9 genes were found in 63% of the fecal isolates. Conclusion The antimicrobial resistant profile found in this study is alarming and poses a great threat to public health. Apparently an increased antimicrobial resistance to the extensively used cephalosporins is affecting an optimal drug therapy for patients. In addition, the presence of catheters, prolonged duration of stay in the hospital and poor hygienic conditions due to infrequent urination of the patient can lead to an additional vulnerability. Therefore continuous surveillance and rational use of antibiotics along with effective hygienic measures are urgently recommended in such settings

The human SIN3B corepressor forms a nucleolar complex with leukemia-associated ETO homologues-2

5) in combination with hSIN3B (S) as described in Materials and Methods. IP and Western were performed with α-V5 (V5 specific) and α-S (hSIN3B specific). () ETO and MTG16 co-precipitated hSIN3B (Lanes 1 and 7 of A respectively). Reciprocal experiments showed that hSIN3B co-precipitated ETO (Lanes 2 and 8 of B respectively). Arrowhead shows the position of ETO in A and MTGR1 in B. MTGR1 failed to co-precipitate hSIN3B (Lane 4 of A). The reciprocal experiment showed that hSIN3B did not precipitate MTGR1 (Lane 5 of B). Lower panels in A and B show input of hSIN3B and ETO homologue in 2% of IP lysate. The positions of the molecular weight markers are indicated at the left.<p><b>Copyright information:</b></p><p>Taken from "The human SIN3B corepressor forms a nucleolar complex with leukemia-associated ETO homologues"</p><p>http://www.biomedcentral.com/1471-2199/9/8</p><p>BMC Molecular Biology 2008;9():8-8.</p><p>Published online 19 Jan 2008</p><p>PMCID:PMC2266940.</p><p></p