Minimally Invasive Skin Transcriptome Extraction Using a Dermal Biomarker Patch.

INTRODUCTION: Advances in the scientific understanding of the skin and characteristic genomic dermal signatures continue to develop rapidly. Nonetheless, skin diagnosis remains predicated on a subjective visual examination, frequently followed by biopsy and histology. These procedures often are not sufficiently sensitive, and in the case of many inflammatory diseases, biopsies are not justified, creating a situation where high-quality samples can be difficult to obtain. The wealth of molecular information available and the pace at which new data are acquired suggest that methods for minimally invasive biomarker collection could dramatically alter our understanding of skin disease and positively impact treatment paradigms. METHODS: A chemical method was optimized to covalently modify custom dermal patches with single-stranded DNA that could bind to messenger RNA. These patches were applied to ex vivo skin samples and penetration evaluated by histological methods. Patches were then applied to both the skin of normal human subjects (lower arm) as well as lesional skin of psoriasis patients, and the transcriptome captured (N = 7; 33 unique samples). Standard RNA-Seq processing was performed to assess the gene detection rate and assessments made of the reproducibility of the extraction procedure as well as the overlap with matched punch biopsy samples from the same patient. RESULTS: We have developed a dermal biomarker patch (DBP) designed to be minimally invasive and extract the dermal transcriptome. Using this platform, we have demonstrated successful molecular analysis from healthy human skin and psoriatic lesions, replicating the molecular information captured with punch biopsy. CONCLUSION: This DBP enables an unprecedented ability to monitor the molecular "fingerprint" of the skin over time or with various interventions, and generate previously inaccessible rich datasets. Furthermore, use of the DBP could be favored by patients relative to biopsy by limiting pain resulting from biopsy procedures. Given the large dynamic range observed in psoriatic skin, analysis of complex phenotypes is now possible, and the power of machine-learning methods can be brought to bear on dermatologic disease.

Re-Assessing PK/PD Issues for Oral Protein and Peptide Delivery.

Due to a lack of safe and effective oral delivery strategies for most protein and peptide therapeutics, pharmaceutical drug developers have focused on parenteral routes to administer these agents. Recent advances in delivery technologies have now shown clinical validation for a few of these biopharmaceuticals following oral administration. While these initial opportunities have provided more than just a glimmer of hope within the industry, there are important aspects of oral biopharmaceutical delivery that do not completely align with pharmacokinetic (PK) parameters and pharmacodynamics (PD) outcomes that have been learned from parenteral administrations. This commentary examines some of these issues with the goal of presenting a rationale for re-assessing methods, models, and success criteria to better measure oral protein or peptide delivery outcomes related to PK/PD events.

A Novel Fusion of IL-10 Engineered to Traffic across Intestinal Epithelium to Treat Colitis.

IL-10 is a potent anti-inflammatory cytokine capable of suppressing a number of proinflammatory signals associated with intestinal inflammatory diseases, such as ulcerative colitis and Crohn's disease. Clinical use of human IL-10 (hIL-10) has been limited by anemia and thrombocytopenia following systemic injection, side effects that might be eliminated by a gut-restricted distribution. We have identified a transcytosis pathway used by cholix, an exotoxin secreted by nonpandemic forms of the intestinal pathogen Vibrio cholerae A nontoxic fragment of the first 386 aa of cholix was genetically fused to hIL-10 to produce recombinant AMT-101. In vitro and in vivo characterization of AMT-101 showed it to efficiently cross healthy human intestinal epithelium (SMI-100) by a vesicular transcytosis process, activate hIL-10 receptors in an engineered U2OS osteosarcoma cell line, and increase cellular phospho-STAT3 levels in J774.2 mouse macrophage cells. AMT-101 was taken up by inflamed intestinal mucosa and activated pSTAT3 in the lamina propria with limited systemic distribution. AMT-101 administered to healthy mice by oral gavage or to cynomolgus monkeys (nonhuman primates) by colonic spray increased circulating levels of IL-1R antagonist (IL-1Ra). Oral gavage of AMT-101 in two mouse models of induced colitis prevented associated pathological events and plasma cytokine changes. Overall, these studies suggest that AMT-101 can efficiently overcome the epithelial barrier to focus biologically active IL-10 to the intestinal lamina propria.

Evaluating maternity care using national administrative health datasets: how are statistics affected by the quality of data on method of delivery?

BACKGROUND: Information on maternity services is increasingly derived from national administrative health data. We evaluated how statistics on maternity care in England were affected by the completeness and consistency of data on "method of delivery" in a national dataset. METHODS: Singleton deliveries occurring between April 2009 and March 2010 in English NHS trusts were extracted from the Hospital Episode Statistics (HES) database. In HES, method of delivery can be entered twice: 1) as a procedure code in core fields, and 2) in supplementary maternity fields. We examined overall consistency of these data sources at a national level and among individual trusts. The impact of different analysis rules for handling inconsistent data was then examined using three maternity statistics: emergency caesarean section (CS) rate; third/fourth degree tear rate amongst instrumental deliveries, and elective CS rate for breech presentation. RESULTS: We identified 629,049 singleton deliveries. Method of delivery was not entered as a procedure or in the supplementary fields in 0.8% and 12.5% of records, respectively. In 545,594 records containing both data items, method of delivery was coded consistently in 96.3% (kappa = 0.93; p < 0.001). Eleven of 136 NHS trusts had comparatively poor consistency (<92%) suggesting systematic data entry errors. The different analysis rules had a small effect on the statistics at a national level but the effect could be substantial for individual NHS trusts. The elective CS rate for breech was most sensitive to the chosen analysis rule. CONCLUSIONS: Organisational maternity statistics are sensitive to inconsistencies in data on method of delivery, and publications of quality indicators should describe how such data were handled. Overall, method of delivery is coded consistently in English administrative health data.

Risk of placenta previa in second birth after first birth cesarean section: a population-based study and meta-analysis.

OBJECTIVE: To compare the risk of placenta previa at second birth among women who had a cesarean section (CS) at first birth with women who delivered vaginally. METHODS: Retrospective cohort study of 399,674 women who gave birth to a singleton first and second baby between April 2000 and February 2009 in England. Multiple logistic regression was used to adjust the estimates for maternal age, ethnicity, deprivation, placenta previa at first birth, inter-birth interval and pregnancy complications. In addition, we conducted a meta-analysis of the reported results in peer-reviewed articles since 1980. RESULTS: The rate of placenta previa at second birth for women with vaginal first births was 4.4 per 1000 births, compared to 8.7 per 1000 births for women with CS at first birth. After adjustment, CS at first birth remained associated with an increased risk of placenta previa (odds ratio = 1.60; 95% CI 1.44 to 1.76). In the meta-analysis of 37 previously published studies from 21 countries, the overall pooled random effects odds ratio was 2.20 (95% CI 1.96-2.46). Our results from the current study is consistent with those of the meta-analysis as the pooled odds ratio for the six population-based cohort studies that analyzed second births only was 1.51 (95% CI 1.39-1.65). CONCLUSIONS: There is an increased risk of placenta previa in the subsequent pregnancy after CS delivery at first birth, but the risk is lower than previously estimated. Given the placenta previa rate in England and the adjusted effect of previous CS, 359 deliveries by CS at first birth would result in one additional case of placenta previa in the next pregnancy.

Maternal morbidity associated with placenta praevia among women who had elective caesarean section.

OBJECTIVE: Estimates of the increased risk of maternal complications after caesarean section posed by placenta praevia differ between studies and may not reflect current practice. We assess the impact of placenta praevia on maternal complications after elective caesarean section (CS). STUDY DESIGN: We undertook a retrospective cohort study of women who had an elective CS for a singleton at term in the English National Health Service between 1 April 2000 and 28 February 2009 using routine data from the Hospital Episode Statistics database. Multiple logistic regression was used to estimate the effect of placenta praevia on maternal complications after controlling for maternal age, parity, whether a woman had a previous CS, and gestational age. Maternal complications included postpartum haemorrhage, obstetric trauma, blood transfusion and hysterectomy. RESULTS: Among 131,731 women having an elective CS for a singleton, 4,332 (3.3%) women had placenta praevia. Placenta praevia increased the risk of postpartum haemorrhage from 9.7% to 17.5% (adjusted odds ratio (OR) 1.91; 95% CI: 1.74 to 2.09), the risk of blood transfusion from 1.4% to 6.4% (OR 4.39; 3.76 to 5.12), and the risk of hysterectomy from 0.03% to 1% (OR 39.70; 22.42 to 70.30). Previous studies have estimated the rate of hysterectomy among women with placenta praevia to be 5%. CONCLUSION: Placenta praevia remains a risk factor for various maternal complications, although the increased risk of hysterectomy is lower than previously reported.

The timing of elective caesarean delivery between 2000 and 2009 in England.

BACKGROUND: In 2004, the National Institute for Clinical Excellence (NICE) recommended that an elective caesarean section for an uncomplicated pregnancy should not be carried out before 39 completed weeks due to increased risk of respiratory morbidity in newborns. We describe the trends and variation across 63 English NHS trusts in the timing of elective caesarean section (CS) for low-risk singleton deliveries. METHODS: We identified elective CS deliveries between 1st April 2000 and 28th February 2009 in English NHS trusts using the Hospital Episode Statistics. We selected women with uncomplicated pregnancies who had an elective CS delivery after 34 completed weeks of gestation, and analysed the trends and the trust-level variation in the timing of elective CS. The impact of the NICE guidance on the monthly rate of elective CS deliveries performed after 39 weeks was estimated using an interrupted time-series design with autoregressive integrated moving average (ARIMA). RESULTS: There were 118,456 elective CS deliveries at the 63 NHS trusts. The overall proportion of elective CS deliveries done after 39 completed weeks steadily increased from 39% in 2000/01 to 63% in 2008/09. The proportions rose from 43% to 67% for women with breech presentation and from 35% to 62% for women with a previous CS. There was significant variation across NHS trusts in each year; in 2008/09, with the proportions of elective CS done after 39 weeks ranging from 28% to 89% (Inter-quartile range limits: 54% to 72%). We found a small but statistically significant increase in the proportion immediately after the publication of the NICE guidance, but its rate of growth rate declined slightly thereafter. CONCLUSIONS: NHS trusts in our study have responded to the new evidence on the benefits of delaying elective CS to after 39 weeks gestation. However, substantial differences between NHS trusts remain, which indicates there is room for further improvement. We suggest that maternity services and commissioners adopt the "timing of elective caesarean" as a quality indicator to support clinical practice.

Influenza A H1N1 2009 (Swine Flu) and Pregnancy.

The Influenza A H1N1 pandemic (A H1N1) occurred between June 2009 and August 2010. Although the pandemic is now over, the virus has emerged as the predominant strain in the current seasonal influenza phase in the northern hemisphere. The A H1N1 influenza is a novel strain of the influenza A virus and is widely known as swine flu. The virus contains a mixture of genetic material from human, pig and bird flu virus. It is a new variety of flu which people have not had much immunity to. Much has been learnt from the Pandemic of 2009/2010 but the messages about vaccination and treatment seem to be taken slowly by the clinical profession. Most people affected by the virus, including pregnant women, suffer a mild viral illness, and make a full recovery. The median duration of illness is around seven days. This influenza typically affects the younger age group i.e. from the ages of 5-65 years. Current experience shows that the age group experiencing increased morbidity and mortality rates are in those under 65 years of age. Pregnant women, because of their altered immunity and physiological adaptations, are at higher risk of developing pulmonary complications, especially in the second and third trimesters. In the United Kingdom, twelve maternal deaths were reported to be associated with the H1N1 virus during the pandemic and clear avoidable factors were identified (Modder, Review of Maternal Deaths in the UK related to A H1N1 2009 influenza (CMACE). www.cmace.org.uk, 2010). The pregnancy outcomes were also poor for women who were affected by the virus with a fivefold increase in the perinatal mortality rate and threefold increase in the preterm delivery rate (Yates et al. Health Technol Assess 14(34):109-182, 2010). There continues to be a low uptake of the flu vaccine and commencement of antiviral treatment for pregnant women.

Variation in rates of caesarean section among English NHS trusts after accounting for maternal and clinical risk: cross sectional study.

OBJECTIVE: To determine whether the variation in unadjusted rates of caesarean section derived from routine data in NHS trusts in England can be explained by maternal characteristics and clinical risk factors. DESIGN: A cross sectional analysis using routinely collected hospital episode statistics was performed. A multiple logistic regression model was used to estimate the likelihood of women having a caesarean section given their maternal characteristics (age, ethnicity, parity, and socioeconomic deprivation) and clinical risk factors (previous caesarean section, breech presentation, and fetal distress). Adjusted rates of caesarean section for each NHS trust were produced from this model. SETTING: 146 English NHS trusts. Population Women aged between 15 and 44 years with a singleton birth between 1 January and 31 December 2008. MAIN OUTCOME MEASURE: Rate of caesarean sections per 100 births (live or stillborn). RESULTS: Among 620 604 singleton births, 147 726 (23.8%) were delivered by caesarean section. Women were more likely to have a caesarean section if they had had one previously (70.8%) or had a baby with breech presentation (89.8%). Unadjusted rates of caesarean section among the NHS trusts ranged from 13.6% to 31.9%. Trusts differed in their patient populations, but adjusted rates still ranged from 14.9% to 32.1%. Rates of emergency caesarean section varied between trusts more than rates of elective caesarean section. CONCLUSION: Characteristics of women delivering at NHS trusts differ, and comparing unadjusted rates of caesarean section should be avoided. Adjusted rates of caesarean section still vary considerably and attempts to reduce this variation should examine issues linked to emergency caesarean section.

Preventing infective complications relating to induced abortion.

Infective complications following induced abortions are still a common cause of morbidity and mortality. This review focusses on defining the strategies to improve care of women seeking an induced abortion and to reduce infective complications. We have considered the evidence for screening and cost-effectiveness for antibiotic prophylaxis. Current evidence suggests that treating all women with prophylactic antibiotics in preference to screening and treating is the most cost-effective way of reducing infective complications following induced abortions. The final strategy to prevent infective complications should be individualized for each region/area depending on the prevalence of organisms causing pelvic infections and the resources available.

Modulation of chondrocyte phenotype for tissue engineering by designing the biologic-polymer carrier interface.

Therapeutic strategies based on cell and tissue engineering can be advanced by developing material substrates that effectively interrogate the biological compartment, with or without the complimentary local release of growth factors. Poly(ether ester) segmented copolymers were engineered as model material systems to elucidate the interfacial molecular events that govern the function of adhered cells. Surface chemistry was modulated by varying poly(ethylene glycol) (PEG) length and mole fraction with poly(butylene terephthalate) (PBT), leading to differential competitive protein adsorption of fibronectin and vitronectin from serum and consequently to different cell attachment modes. Adhesion within the hydrogel-like milieu of longer surface PEG was mediated via binding to the CD44 transmembrane receptor, rather than the RGD-integrin mechanism, whereas greater substrate-bound fibronectin resulted in cell adhesion via integrins. These adhesion modalities differentially impacted morphological cell phenotype (spread or spheroid) and the subsequent expression of mRNA transcripts (collagen types II, I) characteristic of phenotypically differentiated or dedifferentiated chondrocytes, respectively. These results demonstrate that materials can be designed to directly elicit the membrane bound receptor apparatus desired for downstream cellular response, without requiring exogenous biological growth factors to enable differentiated potential.

Evaluation of chondrogenesis within PEGT: PBT scaffolds with high PEG content.

Porous poly(ethylene glycol) terephthalate:poly (butylene terephthalate) (PEGT:PBT) scaffolds with high PEG molecular weight (1000 g/mole) and PEGT content (60%) were fabricated using two different processes-paraffin templating and compression molding-for cartilage engineering applications. This polymer composition has previously been shown to enable chondrocyte adhesion and maintain differentiated phenotype in 2D monolayer culture. The influence of 3D polymer scaffold processing on the formation of cartilaginous tissue was studied by seeding primary immature bovine chondrocytes within cylindrical scaffolds in mixed flask reactors for 3 days, followed by cultivation in culture plates for a total of 10 or 24 days. Tissue-polymer constructs were evaluated morphologically by SEM and histology, and quantitatively for cellularity, total collagen, and glycosaminoglycan content, all of which remained statistically equivalent for each time point tested, irrespective of fabrication method. These data demonstrate that the polymers engineered for this study were able to support chondrogenesis independent of scaffold fabrication process, with the influence of pore architecture lessened by the highly hydrated scaffold microenvironments induced by high PEG content.

Tissue engineering of bovine articular cartilage within porous poly(ether ester) copolymer scaffolds with different structures.

The potential of porous poly(ether ester) scaffolds made from poly(ethylene glycol) terephthalate: poly(butylene terephthalate) (PEGT:PBT) block copolymers produced by various methods to enable cartilaginous tissue formation in vitro was studied. Scaffolds were fabricated by two different processes: paraffin templating (PT) and compression molding (CM). To determine whether PEGT:PBT scaffolds are able to support chondrogenesis, primary bovine chondrocytes were seeded within cylindrical scaffolds under dynamic seeding conditions. On day 3, constructs were transferred to six-well plates and evaluated for glycosaminoglycan (GAG) distribution (3, 10, and 24 days), type II collagen distribution, cellularity, and total collagen and GAG content (10 and 24 days). It was observed that better cell distribution during infiltration within PT scaffolds allowed greater chondrogenesis, and at later time points, than in CM scaffolds. The amount of GAG remained constant for all groups from 10 to 24 days, whereas collagen content increased significantly. These data suggest that PEGT:PBT scaffolds are suitable for cartilage tissue engineering, with the PT process enabling greater chondrogenesis than CM.

Biomaterial microarrays: rapid, microscale screening of polymer-cell interaction.

The identification of biomaterials that induce optimal gene expression patterns and allow for appropriate levels of cellular attachment is of central importance in tissue engineering and cell therapy. Herein, we describe the creation of cell-compatible, biomaterial microarrays, that allow rapid, microscale testing of biomaterial interactions with cells. As proof of principle, we simultaneously characterized over 3456 human mesenchymal stem cell (hMSC)-biomaterial composite interactions, and describe preliminary studies on the utility of these arrays with a neural stem cell line (NSC), and primary articular chondrocytes.

Adhesion-mediated signal transduction in human articular chondrocytes: the influence of biomaterial chemistry and tenascin-C.

Chondrocyte 'dedifferentiation' involves the switching of the cell phenotype to one that no longer secretes extracellular matrix found in normal cartilage and occurs frequently during chondrocyte expansion in culture. It is also characterized by the differential expression of receptors and intracellular proteins that are involved in signal transduction pathways, including those associated with cell shape and actin microfilament organization. The objective of this study was to examine the modulation of chondrocyte phenotype by cultivation on polymer substrates containing poly(ethylene glycol) (PEG). We observed differential arrangement of actin organization in articular chondrocytes, depending on PEG length. When cultivated on 300 g/mol PEG substrates at day 19, chondrocytes had lost intracellular markers characteristic of the differentiated phenotype, including type II collagen and protein kinase C (PKC). On these surfaces, chondrocytes also expressed focal adhesion and signaling proteins indicative of cell attachment, spreading, and FA turnover, including RhoA, focal adhesion kinase, and vinculin. The switch to a dedifferentiated chondrocyte phenotype correlated with integrin expression. Conversely, the expression of CD44 receptors coincided with chondrogenic characteristics, suggesting that binding via these receptors could play a role in maintaining the differentiated phenotype on such substrates. These effects can be similar to those of compounds that interfere in intracellular signaling pathways and can be utilized to engineer cellular response.

Tension free vaginal tape: a procedure for all ages.

OBJECTIVE: To assess the success and complications of the tension free vaginal tape (TVT) procedure in different age groups. PATIENTS AND METHODS: This prospective long-term study of 179 consecutive cases of urodynamically confirmed urinary incontinence that had had the TVT procedure was conducted from March 1999 to December 2002 at a District General Hospital. To assess whether outcome was influenced by the patient's age, the patients were divided into three age groups: group A (30-49 years old), group B (50-69 years old) and group C (70-90 years old). Operative details and early and late complications were recorded, and patients were followed up with clinic visits at 6 weeks and 6 months and a quality of life questionnaire was completed at 1 year. RESULTS: Of the 179 patients included in the study, 53 (29.6%) were in group A, 91 (50.8%) in group B and 35 (19.5%) in group C. The subjective cure rate for the patients was 84.9%, 81.3% and 85.3% in groups A, B and C, respectively. A significant improvement in symptoms was reported by 3.8%, 14.3% and 8.6% women, respectively. The failure rate was 11.3%, 4.4% and 5.7%, respectively. The intraoperative complication rate was 5.6%. The overall postoperative complication rate was 29.6%. A total of 86.2% of the patients were treated as day cases. Patients who had intraoperative complications or initial voiding difficulties (i.e. those patients requiring "in/out" catheterization before spontaneous voiding was established) were in hospital for 1-2 days. CONCLUSIONS: Our data showed better subjective cure rates and substantial improvement rates without any significant increase in intraoperative complications with increasing age. Postoperative complications of urgency and vaginal wall erosion were more common in the older aged patients but were easily resolved. Hospital stay and recovery period were short, making TVT a suitable procedure for all ages.

Tissue engineering of fish skin: behavior of fish cells on poly(ethylene glycol terephthalate)/poly(butylene terephthalate) copolymers in relation to the composition of the polymer substrate as an initial step in constructing a robotic/living tissue hybrid.

This study presents the development of a biosynthetic fish skin to be used on aquatic robots that can emulate fish. Smoothness of the external surface is desired in improving high propulsive efficiency and maneuvering agility of autonomous underwater vehicles such as the RoboTuna (Triantafyllou, M., and Triantafyllou, G. Sci. Am. 272, 64, 1995). An initial step was to determine the seeding density and select a polymer for the scaffolds. The attachment and proliferation of chinook salmon embryo (CHSE-214) and brown bullhead (BB) cells were studied on different compositions of a poly(ethylene glycol terephthalate) (PEGT) and poly(butylene terephthalate) (PBT) copolymer (Polyactive). Polymer films were used, cast of three different compositions of PEGT/PBT (weight ratios of 55/45, 60/40, and 70/30) and two different molecular masses of PEGT (300 and 1000 Da). When a 55 wt% and a 300-Da molecular mass form of PEGT was used, maximum attachment and proliferation of CHSE-214 and BB cells were achieved. Histological studies and immunostaining indicate the presence of collagen and cytokeratins in the extracellular matrix formed after 14 days of culture. Porous scaffolds of PEGT/PBT copolymers were also used for three-dimensional tissue engineering of fish skin, using BB cells. Overall, our results indicate that fish cells can attach, proliferate, and express fish skin components on dense and porous Polyactive scaffolds.

Current knowledge of risks and benefits of prophylactic oophorectomy at hysterectomy for benign disease in United Kingdom and Republic of Ireland.

OBJECTIVE: To estimate current practice of prophylactic oophorectomy at hysterectomy for benign disease and also define the role of the variables considered prior to making the decision of prophylactic oophorectomy among consultant obstetricians and gynaecologists in UK and Republic of Ireland. DESIGN: A postal questionnaire was sent to all 1536 practising consultant gynaecologists in UK and Republic of Ireland. METHODS AND MAIN OUTCOMES MEASURED: Of the 809 replies received, 21% of the respondents routinely performed prophylactic oophorectomy. There was a wide regional variation; 40% consultants in Wales considered prophylactic oophorectomy compared to 16% in the Northern and Yorkshire regions. Consultants with a special interest with a more surgical bias were more likely to consider performing prophylactic oophorectomy. The majority of respondents estimated that prophylactic oophorectomy would reduce risk of ovarian cancer by up to 5%, and that following hysterectomy and ovarian conservation there was an increased risk of premature menopause. CONCLUSION: There is a large variation and uncertainty in the practice of prophylactic oophorectomy in UK and Ireland.