E6 and E7 from Beta Hpv38 Cooperate with Ultraviolet Light in the Development of Actinic Keratosis-Like Lesions and Squamous Cell Carcinoma in Mice

Cutaneous beta human papillomavirus (HPV) types appear to be involved in the development of non-melanoma skin cancer (NMSC); however, it is not entirely clear whether they play a direct role. We have previously shown that E6 and E7 oncoproteins from the beta HPV type 38 display transforming activities in several experimental models. To evaluate the possible contribution of HPV38 in a proliferative tissue compartment during carcinogenesis, we generated a new transgenic mouse model (Tg) where HPV38 E6 and E7 are expressed in the undifferentiated basal layer of epithelia under the control of the Keratin 14 (K14) promoter. Viral oncogene expression led to increased cellular proliferation in the epidermis of the Tg animals in comparison to the wild-type littermates. Although no spontaneous formation of tumours was observed during the lifespan of the K14 HPV38 E6/E7-Tg mice, they were highly susceptible to 7,12-dimethylbenz(a)anthracene (DMBA)/12-0-tetradecanoylphorbol-13-acetate (TPA) two-stage chemical carcinogenesis. In addition, when animals were exposed to ultraviolet light (UV) irradiation, we observed that accumulation of p21WAF1 and cell-cycle arrest were significantly alleviated in the skin of Tg mice as compared to wild-type controls. Most importantly, chronic UV irradiation of Tg mice induced the development of actinic keratosis-like lesions, which are considered in humans as precursors of squamous cell carcinomas (SCC), and subsequently of SCC in a significant proportion of the animals. In contrast, wild-type animals subjected to identical treatments did not develop any type of skin lesions. Thus, the oncoproteins E6 and E7 from beta HPV38 significantly contribute to SCC development in the skin rendering keratinocytes more susceptible to UV-induced carcinogenesis

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

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

p21^WAF1 and Ki-67 levels in the skin of wild-type and K14 HPV38 E6/E7-Tg mice after UVB irradiation.

<p>Wild-type and Tg animals were irradiated up to 5 times as described in Materials and <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002125#s4" target="_blank">Methods</a>. 24 hours after the last irradiation, mice were sacrificed and skin tissue was analyzed by immuno-histochemistry. (A) Representative Ki-67 and p21^WAF1 immunostainings of skin from wild-type and Tg mice non-exposed (0×) or four time (4×) exposed to UVB. (B) Quantification of p21^WAF1 and Ki-67-positive cells in skin of wild-type and Tg mice before and after UVB irradiation. The percentage of p21^WAF1 and Ki-67-positive cells in the epidermis was determined as described in the legend of <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002125#ppat-1002125-g003" target="_blank">Figure 3</a>. The differences between the percentages of p21^WAF1 or Ki-67-positive cells in the HPV38 E6/E7 Tg mice (lines 183 and 187) versus the FVB/N non-Tg mice are statistically significant (* = <i>p</i><0.05, ** = <i>p</i><0,001) as determined by Student's t-test.</p

HPV38 E6 and E7 expression in Tg mice.

<p>(A) Schematic representation of the K14-HPV38 E6/E7 construct. (B) HPV38 E6 and E7 transcripts are differentially expressed in the epithelia of the two hemizygous Tg mouse lines 183, and 187. Total RNA was extracted from the ear, the skin, tongue, esophagus, and liver. After preparation of cDNA, E6 and E7 expression was determined by RT-qPCR and normalized towards the expression level of GAPDH. The data shown in the Figures are the means ±SD of three independent experiments. In each experiment the 187 ear data is set to 1 and the other values are consequently resized.</p

Tumour burden in wild-type and K14 HPV38 E6/E7-Tg animals upon UVB irradiation.

<p>(A) Schematic diagram of the experimental procedure of long-term UVB irradiation. (B) Representative pictures of dorsal skin from wild-type FVB/N and HPV-Tg mice exposed to UVB light for 24 weeks. (C) Representative pictures of HE-stained actinic keratosis affected epidermis (AK) from K14 HPV38 E6/E7-Tg mouse lines 183 and 187 after 24 weeks of irradiation (original magnification 10×). Magnified areas are shown in the right panels. (D) Representative pictures of dorsal skin from wild-type FVB/N and HPV-Tg mice exposed to UVB light for 29 weeks. (E) Percentage of animals with skin SCC in wild-type and Tg cohorts of lines 183 and 187. Tumour formation was monitored each week until the end of the experiment in week 30 after start of treatment, and confirmed by histological analyses after sacrifice of the animals. The difference between the curves of control and transgenic mice is statistically significant (<i>p</i><0,0001 determined by logrank test for group data). (F) Representative pictures of HE-stained of SCC sections (SCC) from K14 HPV38 E6/E7-Tg mouse lines 183 and 187 after 30 weeks of UVB irradiation (original magnification 5×). Magnified areas are shown in the right panels. (G) Representative pictures of HE-stained epidermis from wild-type FVB/N mice after 30 weeks of UVB irradiation (original magnification 10×). Magnified area is shown in the right panel.</p

Histological analysis of skin specimens from wild-type FVB/N and Tg mouse lines.

<p>Representative pictures (original magnification 40×) of HE-stained sections of paraffin-embedded tissues are shown: (A) ear (left panel) and dorsal skin (right panel) of wild-type FVB/N and Tg mice of the lines 183, and 187. (B) Dysplastic ear skin of K14 HPV38 E6/E7-Tg mice.</p