Protein spot arrays on graphene oxide coatings for efficient single-cell capture.

Biomedical applications such as cell screening or cell-cell interaction studies require placement and adhesion of cells on surfaces with controlled numbers and location. In particular, single-cell arraying and positioning has come into focus as a basis of such applications. An ideal substrate would combine biocompatibility with favorable attributes such as pattern stability and easy processing. Here, we present a simple yet effective approach to single-cell arraying based on a graphene oxide (GO) surface carrying protein (fibronectin) microarrays to define cell adhesion points. These capture NIH-3T3 cells, resulting in cell arrays, which are benchmarked against analogous arrays on silanized glass samples. We reveal significant improvement in cell-capture performance by the GO coating with regards to overall cell adhesion and single-cell feature occupancy. This overall improvement of cell-arraying combined with retained transparency of substrate for microscopy and good biocompatibility makes this graphene-based approach attractive for single-cell experiments.

High variability in anatomic patterns of cutaneous photodamage: a population-based study.

BACKGROUND: Skin cancer is strongly associated with photodamaged skin, but body sites are often referred to as 'exposed' or 'unexposed' to sun without recognizing extent of site-specific variation. OBJECTIVES: To assess whole-body patterns of photodamage in an Australian population. METHODS: A random sample of adult residents of Queensland underwent imaging across 10 body sites. Photodamage was graded from images using an ordinal photonumeric scale. We used cluster analysis to identify whole-body photodamage patterns and prevalence proportion ratios (PPRs) to assess associated factors. RESULTS: Of 190 adults (median age 52; 58% males), 58% showed severe or moderate-to-severe photodamage on most body sites. A higher proportion of woman had severe photodamage on the arms (upper: P = 0.002, lower: P = 0.034). A higher proportion of men had moderate or severe photodamage on the lower back (P = 0.004). We identified four photodamage patterns: 'severe general' (n = 24, 13%), 'moderate-severe general' (n = 86, 45%), 'moderate-severe v-neck' (n = 40, 21%) and 'mild-moderate upper body' (n = 12, 6%). All participants with 'severe-general' photodamage were >50 years and more likely to have past skin cancer (PPR: 2.54, 95% CI: 1.44-4.49) than those with 'moderate-severe v-neck' photodamage. Those with 'moderate-severe general' photodamage showed similar associations and were more likely female (PPR: 1.33, 95% CI: 1.04-1.69). Past or current smoking was associated with having higher levels of photodamage, with no smokers in those with 'mild-moderate upper body' photodamage. CONCLUSIONS: Moderate-to-severe photodamage across much of the body is common in Queensland adults and associated with age, sex, past skin cancer and smoking. Assuming a universal pattern of site-specific sun exposure could lead to spurious correlations, while accurate and objective assessment of site-specific photodamage can add to understanding of the development of sun-associated skin cancers, in particular site-specific skin carcinogenesis. Additionally, degree of site-specific photodamage has the potential to assist skin cancer screening.

Skin protective and regenerative effects of RM191A, a novel superoxide dismutase mimetic.

Superoxide dismutase (SOD) is known to be protective against oxidative stress-mediated skin dysfunction. Here we explore the potential therapeutic activities of RM191A, a novel SOD mimetic, on skin. RM191A is a water-soluble dimeric copper (Cu2+-Cu3+)-centred polyglycine coordination complex. It displays 10-fold higher superoxide quenching activity compared to SOD as well as significant antioxidant, anti-inflammatory and immunomodulatory activities through beneficial modulation of several significant inflammatory cytokines in vitro and in vivo. We tested the therapeutic potential of RM191A in a topical gel using a human skin explant model and observed that it significantly inhibits UV-induced DNA damage in the epidermis and dermis, including cyclobutane pyrimidine dimers (CPD), 8-oxo-guanine (8-oxoG) and 8-nitroguanine (8NGO). RM191A topical gel is found to be non-toxic, non-teratogenic and readily distributed in the body of mice. Moreover, it significantly accelerates excisional wound healing, reduces 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation and attenuates age-associated oxidative stress in skin, demonstrating both skin regenerative and geroprotective properties of RM191A.

Quantitative detection of viable helminth ova from raw wastewater, human feces, and environmental soil samples using novel PMA-qPCR methods.

In this study, we have evaluated the efficacy of propidium monoazide quantitative polymerase chain reaction (PMA-qPCR) to differentiate between viable and non-viable Ancylostoma caninum ova. The newly developed method was validated using raw wastewater seeded with known numbers of A. caninum ova. Results of this study confirmed that PMA-qPCR has resulted in average of 88 % reduction (P < 0.05) in gene copy numbers for 50 % viable +50 % non-viable when compared with 100 % viable ova. A reduction of 100 % in gene copies was observed for 100 % non-viable ova when compared with 100 % viable ova. Similar reductions (79-80 %) in gene copies were observed for A. caninum ova-seeded raw wastewater samples (n = 18) collected from wastewater treatment plants (WWTPs) A and B. The newly developed PMA-qPCR method was applied to determine the viable ova of different helminths (A. caninum, A. duodenale, Necator americanus and Ascaris lumbricoides) in raw wastewater, human fecal and soil samples. None of the unseeded wastewater samples were positive for the above-mentioned helminths. N. americanus and A. lumbricoides ova were found in unseeded human fecal and soil samples. For the unseeded human fecal samples (1 g), an average gene copy concentration obtained from qPCR and PMA-qPCR was found to be similar (6.8 × 10(5) ± 6.4 × 10(5) and 6.3 × 10(5) ± 4.7 × 10(5)) indicating the presence of viable N. americanus ova. Among the 24 unseeded soil samples tested, only one was positive for A. lumbricoides. The mean gene copy concentration in the positively identified soil sample was 1.0 × 10(5) ± 1.5 × 10(4) (determined by qPCR) compared to 4.9 × 10(4) ± 3.7 × 10(3) (determined by PMA-qPCR). The newly developed PMA-qPCR methods were able to detect viable helminth ova from wastewater and soil samples and could be adapted for health risk assessment.

Perceived Rewards of Nursing Among Christian Nursing Students in Bangalore, India.

Nurses in India face significant challenges and often migrate to practice nursing abroad. Few studies have focused on the rewards of nursing in India. The aim of this study was to illuminate perceived rewards of nursing among Christian student nurses in Bangalore, India. Photovoice, a participatory action methodology was used, and 14 Christian student nurses participated in the study. Thematic interpretation of photographs, journals, critical group dialog sessions, and observational field notes resulted in the identification of two main themes. These themes included intrinsic rewards and lifelong benefits of nursing in India.

Supplementary breast ultrasound screening in Asian women with negative but dense mammograms-a pilot study.

INTRODUCTION: Dense breasts are common in Asian women and they limit the sensitivity of mammography. This study evaluates the performance of supplementary breast ultrasound screening in Asian women with dense mammograms. MATERIALS AND METHODS: The study was approved by the hospital's Institutional Review Board. A prospective clinical trial was performed between September 2002 and November 2004. Asymptomatic Asian women with negative and dense mammograms were offered supplementary ultrasound screening for breast cancer. Ultrasound assessment was categorised as U1 to U4. U1 and U2 cases were recommended routine interval screening mammography. U3 cases were recommended follow-up ultrasound in 6 months and routine interval screening mammography and U4 cases were recommended biopsy. RESULTS: One hundred and forty-one women with mean age of 45.1 years were enrolled into the study. Mean scan time was 13.0 minutes (± 5.6 minutes) for bilateral vs 11.0 minutes (± 1.4 minutes) for unilateral scans. There were 10 patients and 14 patients in the in the U3 and U4 categories, respectively. Two U4 category patients were diagnosed with malignancy-a-6 mm ductal carcinoma-in-situ and a 13- mm invasive ductal carcinoma. The breast cancer detection rate was 1.4%. Sensitivity and specifi city were 100% (2/2) and 88.5% (92/104) respectively. The positive predictive value was 14.3% (2/14) and the negative predictive value was 100% (92/92). CONCLUSION: This pilot study reveals the usefulness of supplementary ultrasound screening in detecting early stage mammographically and clinically occult breast cancers in Asian women with dense breasts. A larger long-term study is, however, needed to assess its feasibility and impact on breast cancer prognosis.

Supplementary breast ultrasound screening in Asian women with negative but dense mammograms-a pilot study

<p><b>INTRODUCTION</b>Dense breasts are common in Asian women and they limit the sensitivity of mammography. This study evaluates the performance of supplementary breast ultrasound screening in Asian women with dense mammograms.</p><p><b>MATERIALS AND METHODS</b>The study was approved by the hospital's Institutional Review Board. A prospective clinical trial was performed between September 2002 and November 2004. Asymptomatic Asian women with negative and dense mammograms were offered supplementary ultrasound screening for breast cancer. Ultrasound assessment was categorised as U1 to U4. U1 and U2 cases were recommended routine interval screening mammography. U3 cases were recommended follow-up ultrasound in 6 months and routine interval screening mammography and U4 cases were recommended biopsy.</p><p><b>RESULTS</b>One hundred and forty-one women with mean age of 45.1 years were enrolled into the study. Mean scan time was 13.0 minutes (± 5.6 minutes) for bilateral vs 11.0 minutes (± 1.4 minutes) for unilateral scans. There were 10 patients and 14 patients in the in the U3 and U4 categories, respectively. Two U4 category patients were diagnosed with malignancy-a-6 mm ductal carcinoma-in-situ and a 13- mm invasive ductal carcinoma. The breast cancer detection rate was 1.4%. Sensitivity and specifi city were 100% (2/2) and 88.5% (92/104) respectively. The positive predictive value was 14.3% (2/14) and the negative predictive value was 100% (92/92).</p><p><b>CONCLUSION</b>This pilot study reveals the usefulness of supplementary ultrasound screening in detecting early stage mammographically and clinically occult breast cancers in Asian women with dense breasts. A larger long-term study is, however, needed to assess its feasibility and impact on breast cancer prognosis.</p>

Endometrial expression of the insulin-like growth factor system during uterine involution in the postpartum dairy cow.

Rapid uterine involution in the postpartum period of dairy cows is important to achieve a short interval to conception. Expression patterns for members of the insulin-like growth factor (IGF) family were determined by in situ hybridisation at day 14+/-0.4 postpartum (n=12 cows) to investigate a potential role for IGFs in modulating uterine involution. Expression in each uterine tissue region was measured as optical density units and data were analysed according to region and horn. IGF-I mRNA was localized to the sub-epithelial stroma (SES) of inter-caruncular and caruncular endometrium. Both IGF-II and IGF-1R expression was detected in the deep endometrial stroma (DES), the caruncular stroma and myometrium. IGFBP-2, IGFBP-4 and IGFBP-6 mRNAs were all localised to the SES of inter-caruncular and caruncular uterine tissue, and in the DES and caruncular stroma, with IGFBP-4 mRNA additionally expressed in myometrium. IGFBP-3 mRNA was only detectable in luminal epithelium. IGFBP-5 mRNA was found in myometrium, inter-caruncular and caruncular SES and caruncular stroma. These data support a role for IGF-I and IGF-II in the extensive tissue remodelling and repair which the postpartum uterus undergoes to return to its non-pregnant state. The differential expression of binding proteins between tissues (IGFBP-3 in epithelium, IGFBP-2, -4, -5 and -6 in stroma and IGFBP-4 and -5 in myometrium) suggest tight control of IGF activity within each compartment. Differential expression of many members of the IGF family between the significantly larger previously gravid horn and the previously non-gravid horn may relate to differences in their rate of tissue remodelling.

Negative energy balance in dairy cows is associated with specific changes in IGF-binding protein expression in the oviduct.

Negative energy balance (NEB) during early lactation in dairy cows leads to an altered metabolic state that has major effects on the production of IGF family members. Low IGF-I concentrations are associated with poor fertility and therefore we aimed to determine whether NEB exerts a direct effect on IGF expression in the postpartum oviduct. Multiparous Holstein cows were allocated to two treatments (each n=6) designed using differential feeding and milking regimes to produce either mild NEB (MNEB) or severe NEB (SNEB). Animals were slaughtered in week 2 of lactation when divergent metabolic profiles were evident. Oviducts were collected for RNA analysis by real-time RT-PCR and in situ hybridisation. Quantitative measures in oviduct gene expression were obtained for all members of the IGF family (IGF-I/II, IGF-binding proteins (IGFBP) 1-6 and receptors for IGF types 1 and 2), insulin A/B, GH, glucocorticoid and oestrogen alpha/beta. Expression of IGFBP-2 and IGFBP-6 (both of which have a high affinity for IGF-II) was decreased in SNEB relative to MNEB (P<0.05). No other gene was altered by NEB, but IGF-II, IGFBP-3, IGFBP-5 and IGFBP-6 all showed differential expression in different regions of the oviduct. These results indicate that, in addition to low circulating IGF-I after calving, NEB may also influence IGF availability in the oviduct indirectly through changes in specific IGFBP expression. It is possible that the predicted increased signalling by IGF-II may perturb embryo development, contributing to the high rates of embryonic mortality in dairy cows.

Influence of negative energy balance on cyclicity and fertility in the high producing dairy cow.

The peripartum period is of critical importance to subsequent health and fertility. Most cows enter a state of negative energy balance (NEB) associated with many metabolic changes which have carry over effects on the resumption and normality of estrous cyclicity and the success of subsequent inseminations. A dataset on 500 lactations explored the relationships between metabolic traits measured before and after calving with fertility. Stepwise multiple regression analysis showed that longer calving to conception intervals were associated with altered profiles of IGF-I, urea and body condition score. These relationships between metabolic profiles and fertility differed between first lactation cows (which are still growing but produce less milk) and mature animals. Early postpartum the liver undergoes extensive biochemical and morphological modifications to adapt to NEB, the uterus is extensively remodeled and must clear bacterial infections, and the ovary must resume ovulatory cycles. RNA isolated from liver and uterine tissues harvested 2 weeks postpartum from cows in mild (MNEB) and severe (SNEB) energy balance was used to screen the Affymetrix 23K bovine microarray. In liver, SNEB resulted in differential expression of key genes involved in lipid catabolism, gluconeogenesis, and the synthesis and stability of IGF-I. This was accompanied by reduced systemic concentrations of IGF-I which is likely to impact on ovarian function and early embryo development. Within endometrium, cows in SNEB showed histological evidence for higher levels of inflammation and the microarray analysis identified groups of differentially expressed genes involved in tissue remodeling and immune response. This may delay uterine repair after calving, likely contributing to the observed reduction in fertility.

Effect of negative energy balance on the insulin-like growth factor system in pre-recruitment ovarian follicles of post partum dairy cows.

Post partum negative energy balance (NEB) in dairy cattle is associated with a delayed return to ovarian cyclicity and reduced fertility. This study compared the IGF system of pre-recruitment ovarian follicles between cows in mild (n = 6) or severe (n = 6) NEB during early lactation. Ovaries were collected in the second week post partum, when circulating concentrations of IGF-I and glucose were lower (P < 0.01) in severe NEB cows. mRNA expression for IGF-II, type 1 IGF receptor (IGF-1R) and IGF-binding proteins (IGFBP)-1 to IGFBP-6 was determined by in situ hybridisation in individual follicles using radiolabelled oligonucleotide probes. Follicles were classified as very small (1-2.5 mm) or small (2.5-5 mm) and healthy or atretic. Relative mRNA concentrations were measured as optical density (OD) units using image analysis. Thecal IGF-II mRNA expression was highest in very small, healthy follicles (P < 0.05). Granulosa cell IGFBP-2 was the only component to change with EB status, with higher mRNA expression in mild compared with severe NEB cows (P < 0.05). IGFBP-1 and IGFBP-3 mRNA expression were undetectable. IGF-1R, IGFBP-4 and IGFBP-5 mRNA expression were not significantly altered by follicle size or health, but IGFBP-5 tended to increase in atretic follicles. The pattern of IGFBP-6 mRNA expression in theca paralleled that of IGF-II mRNA, with higher (P < 0.05) levels in healthy, very small follicles. In conclusion, the reduced expression of IGFBP-2 mRNA in severe NEB cows may alter the bioavailability of circulating IGF-I and locally produced IGF-II to modulate the pre-recruitment stages of follicles required to maintain normal post partum ovarian cyclicity.

Laterality in bovine behavior in an extensive partially suckled herd and an intensive dairy herd.

Cattle exhibit behavioral laterality, but the consistency and correlation between behaviors are unknown. Behavioral laterality was recorded in two herds of contrasting management intensity. The first was a small, extensively managed herd in Brazil, with cows and calves on rangeland, except when removed for handmilking in stalls. The second was a large, intensive British herd, with cows fed mostly indoors and calves removed for individual rearing soon after birth. In herd 1, the side of the body on which the following behaviors were performed was recorded: rumination (rumination), tail waving (tail), tongue protrusion during the initiation of a feeding bout (feeding), hind leg placement when lying (lying), and front leg initiating walking (walking). The distribution of left and right side dominance was normal for all behaviors, with positive correlations between walking and rumination, tail, and feeding, and between lying and rumination. In herd 2, rumination, feeding, and lying behaviors were similarly recorded, as well as parlor side-preference (parlor) and the side of a track chosen when returning to pasture (track). For all behaviors except track, the extent of left- and right-side dominance was not normally distributed, and more cows than expected showed strong laterality on the right or the left side. Parlor and track lateralities were correlated, indicating that cows that entered one side of the parlor also tended to choose the same side of the track. Strong laterality in the intensively managed herd therefore contrasted with that observed in the extensively managed herd and the reasons for such differences in laterality are uncertain.

Measuring the costs and quality of palliative care: a discussion paper.

In an era of ever increasing emphasis on cost-effectiveness of health care, and with the introduction of the internal market, there is now an urgent need for information on the costs and quality of palliative care. This paper highlights the complexities involved in acquiring and measuring such information and discusses methods which were tried out in an exploratory study. The study took the form of in-depth case studies in two hospices, one in the voluntary sector and one in the NHS.

Purchasing palliative care: availability and cost implications.

This paper reports on a survey of the availability of palliative care in Scotland, in the context of the internal market introduced as part of the NHS reforms. It is based on a survey of both the cost and availability of such service, with a discussion of the implications of this information for purchasers of palliative care.

Iodostearic acid in the treatment of advanced gastrointestinal carcinoma.

This phase II non-comparative trial evaluated the efficacy of intramuscular iodostearic acid in the treatment of 30 patients with advanced carcinoma of the gastrointestinal tract. These included 17 patients with colorectal carcinoma, seven with pancreatic carcinoma, four with gastric carcinoma, one with hepatocellular carcinoma and one with cholangiocarcinoma. Iodostearic acid emulsion (2 ml/1.2 g) followed by depomedrone (0.5 ml/10 mg) was injected intramuscularly twice weekly. No serious toxic effects were observed, the main side effects being local pain at the injection site and hot flushes. There were no partial or complete responses. Stable disease was seen in six of 30 patients (20%) at 3 months and three of 30 patients (10%) at 6 months. In conclusion, intramuscular iodostearic acid is relatively non-toxic, however, it appears to be of little value in the treatment of patients with advanced gastrointestinal carcinomas.