Power Doppler Quantification in Assessing Gestational Trophoblastic Neoplasia.

PURPOSE: The FIGO score cannot accurately stratify low-risk gestational trophoblastic neoplasia (GTN) patients who develop chemoresistance to single agent methotrexate chemotherapy. Tumour vascularisation is a key risk factor and its quantification may provide non-invasive way of complementing risk assessment. MATERIALS AND METHODS: 187 FIGO-staged, low-risk GTN patients were prospectively recruited. Power Doppler ultrasound was analysed using a quantification program. Four diagnostic indicators were obtained comprising the number of colour pixels (NCP), mean dB, power Doppler quantification (PDQ), and percentage of colour pixels (%CP). Each indicator performance was assessed to determine if they could distinguish the subset of low-risk patients who became chemoresistant. RESULTS: There were 111 non-resistant and 76 resistant patients. NCP performed best at distinguishing these two groups where the non-resistant group had an average 3435 (±â€Š2060) pixels and the resistant group 6151 (±â€Š3192) pixels (p < 0.001). PDQ and %CP showed significant differences (p < 0.001) but had poorer performance (area under ROC curves were 72 % and 67 % respectively compared with 75 % for NCP). The mean dB index was not significantly different (p = 0.133). CONCLUSION: Power Doppler ultrasound quantification shows potential for non-invasive assessment of tumour vascularity and can distinguish low-risk GTN patients who become chemoresistant from those who have an uncomplicated course with first line treatment.

Inactivation of Rb in stromal fibroblasts promotes epithelial cell invasion.

Stromal-derived growth factors are required for normal epithelial growth but are also implicated in tumour progression. We have observed inactivation of the retinoblastoma protein (Rb), through phosphorylation, in cancer-associated fibroblasts in oro-pharyngeal cancer specimens. Rb is well known for its cell-autonomous effects on cancer initiation and progression; however, cell non-autonomous functions of Rb are not well described. We have identified a cell non-autonomous role of Rb, using three-dimensional cultures, where depletion of Rb in stromal fibroblasts enhances invasive potential of transformed epithelia. In part, this is mediated by upregulation of keratinocyte growth factor (KGF), which is produced by the depleted fibroblasts. KGF drives invasion of epithelial cells through induction of MMP1 expression in an AKT- and Ets2-dependent manner. Our data identify that stromal fibroblasts can alter the invasive behaviour of the epithelium, and we show that altered expression of KGF can mediate these functions.

Genome-wide characterization reveals complex interplay between TP53 and TP63 in response to genotoxic stress.

In response to genotoxic stress the TP53 tumour suppressor activates target gene expression to induce cell cycle arrest or apoptosis depending on the extent of DNA damage. These canonical activities can be repressed by TP63 in normal stratifying epithelia to maintain proliferative capacity or drive proliferation of squamous cell carcinomas, where TP63 is frequently overexpressed/amplified. Here we use ChIP-sequencing, integrated with microarray analysis, to define the genome-wide interplay between TP53 and TP63 in response to genotoxic stress in normal cells. We reveal that TP53 and TP63 bind to overlapping, but distinct cistromes of sites through utilization of distinctive consensus motifs and that TP53 is constitutively bound to a number of sites. We demonstrate that cisplatin and adriamycin elicit distinct effects on TP53 and TP63 binding events, through which TP53 can induce or repress transcription of an extensive network of genes by direct binding and/or modulation of TP63 activity. Collectively, this results in a global TP53-dependent repression of cell cycle progression, mitosis and DNA damage repair concomitant with activation of anti-proliferative and pro-apoptotic canonical target genes. Further analyses reveal that in the absence of genotoxic stress TP63 plays an important role in maintaining expression of DNA repair genes, loss of which results in defective repair.

Mapping human resources for eye health in 21 countries of sub-Saharan Africa: current progress towards VISION 2020.

BACKGROUND: Development of human resources for eye health (HReH) is a major focus of the Global Action Plan 2014 to 2019 to reduce the prevalence of avoidable visual impairment by 25% by the year 2019. The eye health workforce is thought to be much smaller in sub-Saharan Africa than in other regions of the world but data to support this for policy-making is scarce. We collected HReH and cataract surgeries data from 21 countries in sub-Sahara to estimate progress towards key suggested population-based VISION 2020 HReH indicators and cataract surgery rates (CSR) in 2011. METHODS: Routinely collected data on practitioner and surgery numbers in 2011 was requested from national eye care coordinators via electronic questionnaires. Telephone and e-mail discussions were used to determine data collection strategies that fit the national context and to verify reported data quality. Information was collected on six practitioner cadres: ophthalmologists, cataract surgeons, ophthalmic clinical officers, ophthalmic nurses, optometrists and 'mid-level refractionists' and combined with publicly available population data to calculate practitioner to population ratios and CSRs. Associations with development characteristics were conducted using Wilcoxon rank sum tests and Spearman rank correlations. RESULTS: HReH data was not easily available. A minority of countries had achieved the suggested VISION 2020 targets in 2011; five countries for ophthalmologists/cataract surgeons, four for ophthalmic nurses/clinical officers and two for CSR. All countries were below target for optometrists, even when other cadres who perform refractions as a primary duty were considered. The regional (sample) ratio for surgeons (ophthalmologists and cataract surgeons) was 2.9 per million population, 5.5 for ophthalmic clinical officers and nurses, 3.7 for optometrists and other refractionists, and 515 for CSR. A positive correlation between GDP and CSR as well as many practitioner ratios was observed (CSR P = 0.0042, ophthalmologists P = 0.0034, cataract surgeons, ophthalmic nurses and optometrists 0.1 > P > 0.05). CONCLUSIONS: With only a minority of countries in our sample having reached suggested ophthalmic cadre targets and none having reached targets for refractionists in 2011, substantially more targeted investment in HReH may be needed for VISION 2020 aims to be achieved in sub-Saharan Africa.

Trends and implications for achieving VISION 2020 human resources for eye health targets in 16 countries of sub-Saharan Africa by the year 2020.

BACKGROUND: Development of human resources for eye health (HReH) is a major global eye health strategy to reduce the prevalence of avoidable visual impairment by the year 2020. Building on our previous analysis of current progress towards key HReH indicators and cataract surgery rates (CSRs), we predicted future indicator achievement among 16 countries of sub-Saharan Africa by 2020. METHODS: Surgical and HReH data were collected from national eye care programme coordinators on six practitioner cadres: ophthalmologists, cataract surgeons, ophthalmic clinical officers, ophthalmic nurses, optometrists and 'mid-level refractionists' and combined them with publicly available population data to calculate practitioner-to-population ratios and CSRs. Data on workforce entry and exit (2008 to 2010) was used to project practitioner population and CSR growth between 2011 and 2020 in relation to projected growth in the general population. Associations between indicator progress and the presence of a non-physician cataract surgeon cadre were also explored using Wilcoxon rank sum tests and Spearman rank correlations. RESULTS: In our 16-country sample, practitioner per million population ratios are predicted to increase slightly for surgeons (ophthalmologists/cataract surgeons, from 3.1 in 2011 to 3.4 in 2020) and ophthalmic nurses/clinical officers (5.8 to 6.8) but remain low for refractionists (including optometrists, at 3.6 in 2011 and 2020). Among countries that have not already achieved target indicators, however, practitioner growth will be insufficient for any additional countries to reach the surgeon and refractionist targets by year 2020. Without further strategy change and investment, even after 2020, surgeon growth is only expected to sufficiently outpace general population growth to reach the target in one country. For nurses, two additional countries will achieve the target while one will fall below it. In 2011, high surgeon practitioner ratios were associated with high CSR, regardless of the type of surgeon employed. The cataract surgeon workforce is growing proportionately faster than the ophthalmologist. CONCLUSIONS: The HReH workforce is not growing fast enough to achieve global eye health targets in most of the sub-Saharan countries we surveyed by 2020. Countries seeking to make rapid progress to improve CSR could prioritise investment in training new cataract surgeons over ophthalmologists and improving surgical output efficiency.

Genome-wide analysis of p63 binding sites identifies AP-2 factors as co-regulators of epidermal differentiation.

The p63 transcription factor (TP63) is critical in development, growth and differentiation of stratifying epithelia. This is highlighted by the severity of congenital abnormalities caused by TP63 mutations in humans, the dramatic phenotypes in knockout mice and de-regulation of TP63 expression in neoplasia altering the tumour suppressive roles of the TP53 family. In order to define the normal role played by TP63 and provide the basis for better understanding how this network is perturbed in disease, we used chromatin immunoprecipitation combined with massively parallel sequencing (ChIP-seq) to identify >7500 high-confidence TP63-binding regions across the entire genome, in primary human neonatal foreskin keratinocytes (HFKs). Using integrative strategies, we demonstrate that only a subset of these sites are bound by TP53 in response to DNA damage. We identify a role for TP63 in transcriptional regulation of multiple genes genetically linked to cleft palate and identify AP-2alpha (TFAP2A) as a co-regulator of a subset of these genes. We further demonstrate that AP-2gamma (TFAP2C) can bind a subset of these regions and that acute depletion of either TFAP2A or TFAP2C alone is sufficient to reduce terminal differentiation of organotypic epidermal skin equivalents, indicating overlapping physiological functions with TP63.

p63 maintains keratinocyte proliferative capacity through regulation of Skp2-p130 levels.

p63 is a master regulator of proliferation and differentiation in stratifying epithelia, and its expression is frequently altered in carcinogenesis. However, its role in maintaining proliferative capacity remains unclear. Here, we demonstrate that hypoproliferation and loss of differentiation in organotypic raft cultures of primary neonatal human foreskin keratinocytes (HFKs) depleted of the α and ß isoforms of p63 result from p53-p21-mediated accumulation of retinoblastoma (Rb) family member p130. Hypoproliferation in p63-depleted HFKs can be rescued by depletion of p53, p21(CIP1) or p130. Furthermore, we identified the gene encoding S-phase kinase-associated protein 2 (Skp2), the recognition component of the SCF(Skp2) E3 ubiquitin ligase, as a novel target of p63, potentially influencing p130 levels. Expression of Skp2 is maintained by p63 binding to a site in intron 2 and mRNA levels are downregulated in p63-depleted cells. Hypoproliferation in p63-depleted cells can be restored by re-expression of Skp2. Taken together, these results indicate that p63 plays a multifaceted role in maintaining proliferation in the mature regenerating epidermis, in addition to being required for differentiation.

Risk of trachoma in a SAFE intervention area.

To determine prevalence and risk factors of trachoma in communities receiving intervention with the SAFE strategy (surgery, antibiotic, face washing, environmental hygiene), a cross-sectional trachoma survey was undertaken in 2006 in the Enemor district of southern Ethiopia where the SAFE program has been implemented for over five years. A sample of 374 household heads and 2,080 individuals were interviewed and examined for trachoma using an established trachoma grading system of the World Health Organization. The most prominent risk factors were identified with logistic regression analysis. Among individuals >14 years of age, the prevalence of trichiasis was 9.04 % [confidence interval (CI) 7.4-10.6]. People >40 years of age [odds ratio (OR) 1.7; CI 1.2-2.7), women (OR 2.2; CI 1.1-4.3), and illiterates (OR 3; CI 1.4-6.8) had increased risk of trichiasis. Coverage of surgical and antibiotic services was 46 and 85.5 %, respectively. Prevalence of active follicular trachoma (TF) in children aged 1-9 years was 33.1 % (CI 29.4-37.1). Unclean faces (OR 5.9; CI 4.3-8.3) and not being in school (OR 2.1; CI 1.3-3.3) were significantly associated with TF. Clean faces were observed in 56.1 % of children and improved with age and schooling (P < 0.001, Chi-squared test). Household latrine use (74.4 %) was associated with knowledge about SAFE and economic level (P ≤ 0.004, Chi-squared tests). Elderly illiterate women remain at risk of becoming blind from trachoma even in intervention areas. Trachoma particularly affects children without clean faces or opportunity for schooling. Provision of SAFE services with high coverage should be sustained in trachoma-hyperendemic areas.

Evidence for alteration of EZH2, BMI1, and KDM6A and epigenetic reprogramming in human papillomavirus type 16 E6/E7-expressing keratinocytes.

A number of epigenetic alterations occur in both the virus and host cellular genomes during human papillomavirus (HPV)-associated carcinogenesis, and investigations of such alterations, including changes in chromatin proteins and histone modifications, have the potential to lead to therapeutic epigenetic reversion. We report here that transformed HPV16 E6/E7-expressing primary human foreskin keratinocytes (HFKs) (E6/E7 cells) demonstrate increased expression of the PRC2 methyltransferase EZH2 at both the mRNA and protein levels but do not exhibit the expected increase in trimethylated H3K27 (H3K27me3) compared to normal keratinocytes. In contrast, these cells show a reduction in global H3K27me3 levels in vitro, as well as upregulation of the KDM6A demethylase. We further show for the first time that transformation with the HPV16 E6 and E7 oncogenes also results in an increase in phosphorylated EZH2 serine 21 (P-EZH2-Ser21), mediated by active Akt, and in a downregulation of the PRC1 protein BMI1 in these cells. High-grade squamous cervical intraepithelial lesions also showed a loss of H3K27me3 in the presence of increased expression of EZH2. Correlating with the loss of H3K27me3, E6/E7 cells exhibited derepression of specific EZH2-, KMD6A-, and BMI1-targeted HOX genes. These results suggest that the observed reduction in H3K27me3 may be due to a combination of reduced activities/levels of specific polycomb proteins and increases in demethylases. The dysregulation of multiple chromatin proteins resulting in the loss of global H3K27me3 and the transcriptional reprogramming in HPV16 E6/E7-infected cells could provide an epigenetic signature associated with risk and/or progression of HPV16-associated cancers, as well as the potential for epigenetic reversion in the future.

Emergency contraception after sexual assault: changes in provision from 2004 to 2009.

OBJECTIVE: A random sample (20%) of U.S. and territorial emergency departments were surveyed in 2004 and again in 2009 to obtain information about provision and counseling of emergency contraception (EC) to sexual assault victims. STUDY DESIGN: A representative sample of 20% of hospitals, stratified by state/ territory was prepared from the American Hospital Association list in order to conduct a 13-question telephone survey. Questions included (1) "Is there a written protocol for counseling about EC for sexual assault victims?" (2) "Are sexual assault victims at risk of pregnancy counseled about EC?" and (3) "Are sexual assault victims at risk of pregnancy provided EC?" A cross-sectional prevalence survey was administered in 2004 and 2009. RESULTS: Provision of EC has changed very little from 2004 to 2009 (63% vs. 64%, respectively). Provision varies by number of victims treated, region of country and status of state legislation. CONCLUSION: Prophylaxis against possible pregnancy is an important part of sexual assault treatment and should be maximized. EC provision for sexual assault victims in emergency departments has not greatly increased over time and does not reflect regulatory changes in accessibility. Prophylaxes against sexually transmitted infections and pregnancy are handled differently for sexual assault victims, reflecting distinct separation of sexual and reproductive health in clinical practice.

Ophthalmic Telesurgery with a Low-Cost Smartphone Video System for Surgeon Self-Reflection and Remote Synchronous Consultation: A Qualitative and Quantitative Study.

Summary: More than a third of the global burden of blindness is due to cataracts, yet cataract surgery is one of the most cost-effective surgical treatments in medicine. Poor surgical outcomes in many settings remain a major challenge, raising concerns about the quality and efficacy of surgical training. Reflective learning from video recordings of a trainees' surgical performance has a high educational impact and is available routinely for surgical training within high-resource institutions. However, the prohibitive cost and limited portability of current surgical video recording systems make its use problematic in low-resource settings and outreach environments. Objective: The study's aim was to evaluate the potential of smartphone-captured surgical videos for surgeon learning via self-recording and self-review as well as the potential to support live telesurgical consultation. Methodology: A quantitative and qualitative methodology was used to explore and describe the utility and acceptance of smartphone videos in two training facilities in Nepal. Twenty surgeries were recorded on the smartphone for surgeon self-review, to assess image quality, and its application to measure performance against the International Council of Ophthalmology (ICO) Ophthalmology Surgical Competency Assessment Rubrics (OSCAR) SICS Rubric. The same system was used to transmit 15 different surgeries live via Skype from Nepal to an ophthalmologist surgical trainer in South Africa to evaluate the feasibility of live consultation. Findings: Overall video quality was described as high in 65% and moderate in 35% for the videos recorded for self-review. In the surgeries streamed via Skype, quality was described as high in 92.9% and moderate in 7.1%. There were no instances where the video quality was described as poor. The video quality was good enough that the surgeons could measure against ICO-OSCAR rubric in all cases. Discussion: The video quality of smartphone-captured surgical videos was found to be high and gained acceptance for reflective teaching and learning purposes. The extended telesurgical potential and portability of the smartphone enables use across many settings. More studies over a longer period are needed to determine how they can support training and learning in cataract surgery.