Chromatin-modifying agents convert fibroblasts to OCT4+ and VEGFR-2+ capillary tube-forming cells

Rationale The human epigenome is plastic. The goal of this study was to address if fibroblast cells can be epigenetically modified to promote neovessel formation. Methods and results Here, we used highly abundant human adult dermal fibroblast cells (hADFCs) that were treated with the chromatin-modifying agents 5-aza-2\u27-deoxycytidine and trichostatin A, and subsequently subjected to differentiation by activating Wnt signaling. Our results show that these epigenetically modified hADFCs increasingly expressed β-catenin, pluripotency factor octamer-binding transcription factor-4 (OCT4, also known as POU5F1), and endothelial cell (EC) marker called vascular endothelial growth factor receptor-2 (VEGFR-2, also known as Fetal Liver Kinase-1). In microscopic analysis, β-catenin localized to cell-cell contact points, while OCT4 was found to be localized primarily to the nucleus of these cells. Furthermore, in a chromatin immunoprecipitation experiment, OCT4 bound to the VEGFR-2/FLK1 promoter. Finally, these modified hADFCs also transduced Wnt signaling. Importantly, on a two-dimensional (2D) gel substrate, a subset of the converted cells formed vascular network- like structures in the presence of VEGF. Conclusion Chromatin-modifying agents converted hADFCs to OCT4+ and VEGFR-2+ capillary tubeforming cells in a 2D matrix in VEGF-dependent manner

Prevalencija i otpornost na lijekove šiga-toksigenih (STEC), enteropatogenih (EPEC) i enterotoksigenih (ETEC) izolata bakterije Escherichia coli u slobodno držanih jakova s proljevom i bez proljeva u pokrajini West Kameng, Arunachal Pradesh, Indija.

Food producing animals are the major reservoirs of Shiga-toxin producing (STEC) and enteropathogenic Escherichia coli (EPEC). The yak (Poephagus grunniens) is a unique multipurpose bovid, reared by highlanders in the Himalayan region. A total of 67 STEC, 5 EPEC and 22 ETEC strains were isolated from 256 rectal swabs of free ranging yaks from the West Kameng district of Arunachal Pradesh, India. Among the STEC isolates, shiga toxin producing genes stx2 was predominant, followed by stx1. Of all the stx variants, stx1c, stx2d, stx2c, stx2e and stx2f were detected in 23, 11, 2 and 1 isolates, respectively. Further, genes such as eaeA, ehxA, saa, iha and toxB were detected in 16, 35, 28, 10 and 2 isolates, respectively. One of the EPEC isolates possessed a bfpA gene and was categorized as typical EPEC. Among the ETEC isolates, genes such as LT, STa, STb, F41 and K99 were detected in 14, 5, 13, 2 and 2 isolates, respectively. The majority of the STEC, EPEC and ETEC isolates exhibited multi-drug resistance. The study revealed the presence of multi-drug resistant diarrhoea genic E. coli in free ranging yaks from the West Kameng district of Arunachal Pradesh. Moreover, the presence of STEC/EPEC can be a potential public health risk for tribal highlanders residing in close proximity of the reservoir yaks.Životinje koje služe za proizvodnju hrane mogu biti rezervoar šiga-toksigenih (STEC) i enteropatogenih (EPEC) sojeva bakterije Escherichia coli. Jak (Poephagus grunniens) je jedinstveni multipari bovid kojeg uzgajaju gorštaci na Himalaji. Ukupno je bilo izdvojeno 67 STEC sojeva, 5 EPEC sojeva i 22 ETEC soja iz 256 rektalnih obrisaka slobodno držanih jakova na području West Kameng, Arunachal Pradesh, Indija. Među izolatima STEC prevladavao je gen stx2 za prizvodnju šiga-toksina, a slijedio ga je gen stx1. Od svih varijanata gena stx, stx1c bio je dokazan u 23 izolata, stx2d u 11 izolata, stx2c u dva izolata, a stx2e i stx2f u jednog izolata. Nadalje gen eaeA dokazan je u 16 izolata, gen ehxA u 35, gen saa u 28, gen iha u 10 te gen toxBu dva izolata. Jedan od izolata EPEC posjedovao je gen bfpA i svrstan je u tipične EPEC izolate. Gen LT dokazan je u 14 ETEC izolata, gen Sta u pet, gen STb u 13, gen F41 u dva i K99 također u dva izolata. Većina izolata STEC, EPEC i ETEC pokazivala je otpornost na više lijekova. Istraživanje je pokazalo da postoji višestruka otpornost bakterije E. coli izdvojene iz slobodno držanih jakova s proljevom na području West Kameng, Arunachal Pradesh. Povrh toga, prisutnost STEC/EPEC može predstavljati rizik za javno zdravstvo za gorštačka plemena koja dolaze u bliski dodir s jakovima

Prevalencija i otpornost na lijekove šiga-toksigenih (STEC), enteropatogenih (EPEC) i enterotoksigenih (ETEC) izolata bakterije Escherichia coli u slobodno držanih jakova s proljevom i bez proljeva u pokrajini West Kameng, Arunachal Pradesh, Indija.

Food producing animals are the major reservoirs of Shiga-toxin producing (STEC) and enteropathogenic Escherichia coli (EPEC). The yak (Poephagus grunniens) is a unique multipurpose bovid, reared by highlanders in the Himalayan region. A total of 67 STEC, 5 EPEC and 22 ETEC strains were isolated from 256 rectal swabs of free ranging yaks from the West Kameng district of Arunachal Pradesh, India. Among the STEC isolates, shiga toxin producing genes stx2 was predominant, followed by stx1. Of all the stx variants, stx1c, stx2d, stx2c, stx2e and stx2f were detected in 23, 11, 2 and 1 isolates, respectively. Further, genes such as eaeA, ehxA, saa, iha and toxB were detected in 16, 35, 28, 10 and 2 isolates, respectively. One of the EPEC isolates possessed a bfpA gene and was categorized as typical EPEC. Among the ETEC isolates, genes such as LT, STa, STb, F41 and K99 were detected in 14, 5, 13, 2 and 2 isolates, respectively. The majority of the STEC, EPEC and ETEC isolates exhibited multi-drug resistance. The study revealed the presence of multi-drug resistant diarrhoea genic E. coli in free ranging yaks from the West Kameng district of Arunachal Pradesh. Moreover, the presence of STEC/EPEC can be a potential public health risk for tribal highlanders residing in close proximity of the reservoir yaks.Životinje koje služe za proizvodnju hrane mogu biti rezervoar šiga-toksigenih (STEC) i enteropatogenih (EPEC) sojeva bakterije Escherichia coli. Jak (Poephagus grunniens) je jedinstveni multipari bovid kojeg uzgajaju gorštaci na Himalaji. Ukupno je bilo izdvojeno 67 STEC sojeva, 5 EPEC sojeva i 22 ETEC soja iz 256 rektalnih obrisaka slobodno držanih jakova na području West Kameng, Arunachal Pradesh, Indija. Među izolatima STEC prevladavao je gen stx2 za prizvodnju šiga-toksina, a slijedio ga je gen stx1. Od svih varijanata gena stx, stx1c bio je dokazan u 23 izolata, stx2d u 11 izolata, stx2c u dva izolata, a stx2e i stx2f u jednog izolata. Nadalje gen eaeA dokazan je u 16 izolata, gen ehxA u 35, gen saa u 28, gen iha u 10 te gen toxBu dva izolata. Jedan od izolata EPEC posjedovao je gen bfpA i svrstan je u tipične EPEC izolate. Gen LT dokazan je u 14 ETEC izolata, gen Sta u pet, gen STb u 13, gen F41 u dva i K99 također u dva izolata. Većina izolata STEC, EPEC i ETEC pokazivala je otpornost na više lijekova. Istraživanje je pokazalo da postoji višestruka otpornost bakterije E. coli izdvojene iz slobodno držanih jakova s proljevom na području West Kameng, Arunachal Pradesh. Povrh toga, prisutnost STEC/EPEC može predstavljati rizik za javno zdravstvo za gorštačka plemena koja dolaze u bliski dodir s jakovima

Prevalence, associated factors and outcomes of pressure injuries in adult intensive care unit patients: the DecubICUs study

Funder: European Society of Intensive Care Medicine; doi: http://dx.doi.org/10.13039/501100013347Funder: Flemish Society for Critical Care NursesAbstract: Purpose: Intensive care unit (ICU) patients are particularly susceptible to developing pressure injuries. Epidemiologic data is however unavailable. We aimed to provide an international picture of the extent of pressure injuries and factors associated with ICU-acquired pressure injuries in adult ICU patients. Methods: International 1-day point-prevalence study; follow-up for outcome assessment until hospital discharge (maximum 12 weeks). Factors associated with ICU-acquired pressure injury and hospital mortality were assessed by generalised linear mixed-effects regression analysis. Results: Data from 13,254 patients in 1117 ICUs (90 countries) revealed 6747 pressure injuries; 3997 (59.2%) were ICU-acquired. Overall prevalence was 26.6% (95% confidence interval [CI] 25.9–27.3). ICU-acquired prevalence was 16.2% (95% CI 15.6–16.8). Sacrum (37%) and heels (19.5%) were most affected. Factors independently associated with ICU-acquired pressure injuries were older age, male sex, being underweight, emergency surgery, higher Simplified Acute Physiology Score II, Braden score 3 days, comorbidities (chronic obstructive pulmonary disease, immunodeficiency), organ support (renal replacement, mechanical ventilation on ICU admission), and being in a low or lower-middle income-economy. Gradually increasing associations with mortality were identified for increasing severity of pressure injury: stage I (odds ratio [OR] 1.5; 95% CI 1.2–1.8), stage II (OR 1.6; 95% CI 1.4–1.9), and stage III or worse (OR 2.8; 95% CI 2.3–3.3). Conclusion: Pressure injuries are common in adult ICU patients. ICU-acquired pressure injuries are associated with mainly intrinsic factors and mortality. Optimal care standards, increased awareness, appropriate resource allocation, and further research into optimal prevention are pivotal to tackle this important patient safety threat

Prevalence, associated factors and outcomes of pressure injuries in adult intensive care unit patients: the DecubICUs study

Funder: European Society of Intensive Care Medicine; doi: http://dx.doi.org/10.13039/501100013347Funder: Flemish Society for Critical Care NursesAbstract: Purpose: Intensive care unit (ICU) patients are particularly susceptible to developing pressure injuries. Epidemiologic data is however unavailable. We aimed to provide an international picture of the extent of pressure injuries and factors associated with ICU-acquired pressure injuries in adult ICU patients. Methods: International 1-day point-prevalence study; follow-up for outcome assessment until hospital discharge (maximum 12 weeks). Factors associated with ICU-acquired pressure injury and hospital mortality were assessed by generalised linear mixed-effects regression analysis. Results: Data from 13,254 patients in 1117 ICUs (90 countries) revealed 6747 pressure injuries; 3997 (59.2%) were ICU-acquired. Overall prevalence was 26.6% (95% confidence interval [CI] 25.9–27.3). ICU-acquired prevalence was 16.2% (95% CI 15.6–16.8). Sacrum (37%) and heels (19.5%) were most affected. Factors independently associated with ICU-acquired pressure injuries were older age, male sex, being underweight, emergency surgery, higher Simplified Acute Physiology Score II, Braden score 3 days, comorbidities (chronic obstructive pulmonary disease, immunodeficiency), organ support (renal replacement, mechanical ventilation on ICU admission), and being in a low or lower-middle income-economy. Gradually increasing associations with mortality were identified for increasing severity of pressure injury: stage I (odds ratio [OR] 1.5; 95% CI 1.2–1.8), stage II (OR 1.6; 95% CI 1.4–1.9), and stage III or worse (OR 2.8; 95% CI 2.3–3.3). Conclusion: Pressure injuries are common in adult ICU patients. ICU-acquired pressure injuries are associated with mainly intrinsic factors and mortality. Optimal care standards, increased awareness, appropriate resource allocation, and further research into optimal prevention are pivotal to tackle this important patient safety threat

Correction to: Prevalence, associated factors and outcomes of pressure injuries in adult intensive care unit patients: the DecubICUs study

The original version of this article unfortunately contained a mistake

Extension of the fuzzy c means clustering algorithm to fit with the composite graph model for web document representation

Clustering techniques are mostly unsupervised methods that can be used to organize data into groups based on similarities among the individual data items. Fuzzy c-means (FCM) clustering is one of well known unsupervised clustering techniques, which can also be used for unsupervised web document clustering. In this chapter we will introduce a modified method of clustering where the data to be clustered will be represented by graphs instead of vectors or other models. Specifically, we will extend the classical FCM clustering algorithm to work with graphs that represent web documents [1,2,3]. We wish to use graphs because they can allow us to retain information which is often discarded in simpler models

Chromatin-modifying agents convert fibroblasts to OCT4+ and VEGFR-2+ capillary tube-forming cells

<div><p>Rationale</p><p>The human epigenome is plastic. The goal of this study was to address if fibroblast cells can be epigenetically modified to promote neovessel formation.</p><p>Methods and results</p><p>Here, we used highly abundant human adult dermal fibroblast cells (hADFCs) that were treated with the chromatin-modifying agents 5-aza-2'-deoxycytidine and trichostatin A, and subsequently subjected to differentiation by activating Wnt signaling. Our results show that these epigenetically modified hADFCs increasingly expressed β-catenin, pluripotency factor octamer-binding transcription factor-4 (OCT4, also known as POU5F1), and endothelial cell (EC) marker called vascular endothelial growth factor receptor-2 (VEGFR-2, also known as Fetal Liver Kinase-1). In microscopic analysis, β-catenin localized to cell-cell contact points, while OCT4 was found to be localized primarily to the nucleus of these cells. Furthermore, in a chromatin immunoprecipitation experiment, OCT4 bound to the <i>VEGFR-2/FLK1</i> promoter. Finally, these modified hADFCs also transduced Wnt signaling. Importantly, on a two-dimensional (2D) gel substrate, a subset of the converted cells formed vascular network-like structures in the presence of VEGF.</p><p>Conclusion</p><p>Chromatin-modifying agents converted hADFCs to OCT4+ and VEGFR-2+ capillary tube-forming cells in a 2D matrix in VEGF-dependent manner.</p></div

A model of the conversion process of fibroblast cells into tube-forming cells.

<p>hADFCs are mature differentiated cells with highly condensed chromatin containing histones tightly bound to DNAs. However, upon treatment with Aza and TSA for two or three cycles, histone acetylation increases, DNA methylation decreases, the chromatin becomes relaxed, so that the transcription factors could activate the expression of mesodermal-endothelial genes. These events mediate the formation of tube-like structures in 2D Matrigel in a VEGF-dependent manner.</p

Microscopic analysis of β-catenin, OCT4, N-cadherin and vWF in epigenetically modified cells.

<p>hADFCs were plated on coverslips, left untreated or treated with epigenetic modifiers as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0176496#pone.0176496.g001" target="_blank">Fig 1A</a>, and stained with anti-β-catenin (A-D, green) or anti-OCT4 (E-P, green) or anti-vWF (Q-T, red). Representative microscopic images of: <b>A&E</b>) d2 control untreated cells; <b>B&F</b>) d3 cells treated once with Aza + TSA; <b>C&G</b>) d4 cells received two doses of Aza + TSA; <b>D&H</b>) d5, treated with a third dose of Aza + TSA and TDG. White arrowheads indicate β-catenin; magenta arrowheads show nuclear accumulation of OCT4. <b>I-L</b>) d5 cells treated with a third dose of Aza + TSA and TDG were stained with DAPI (blue), TRITC-phalloidin (red), and OCT4 (green). Magenta arrows (<b>L</b>) indicate nuclear localization. <b>M-P</b>) d5 cells treated with a third dose of Aza + TSA and TDG were stained with DAPI (blue), OCT4 (green), and N-cadherin (red). <b>Q-T</b>) d2, d3, d4 and d5 hADFC were stained with DAPI (blue) and vWF (red). Magnification is as shown.</p