Plasmonic nanocone arrays for rapid and detailed cell lysate surface enhanced Raman spectroscopy analysis.

In this work, we develop, fabricate, and characterize a plasmonic nanocone array surface enhanced Raman spectroscopy (SERS) substrate with a uniform enhancement factor on the micron scale for qualitative and quantiative cell and cell lysate analysis. This work demonstrates how SERS substrates can be used as cell-based biosensors given that the enhancement factor of the substrate is sufficient for Raman detection and that the uniformity is high over the applicable surface area. These requirements allow accurate and quantitative comparisons between nonuniform samples under varying biochemical conditions. We apply the developed SERS substrate for Raman measurements and mapping of HeLa cells and cell lysate. This method is used for identification of UV-induced damage and detection of nanomolar concentrations of methylated guanine spiked in cell lysate samples.

Quantitative phase imaging with partially coherent illumination.

In this Letter, we formulate a mathematical model for predicting experimental outcomes in quantitative phase imaging (QPI) when the illumination field is partially spatially coherent. We derive formulae that apply to QPI and discuss expected results for two classes of QPI experiments: common path and traditional interferometry, under varying degrees of spatial coherence. In particular, our results describe the physical relationship between the spatial coherence of the illuminating field and the halo effect, which is well known in phase-contrast microscopy. We performed experiments relevant to this common situation and found that our theory is in excellent agreement with the data. With this new understanding of the effects of spatial coherence, our formulae offer an avenue for removing halo artifacts from phase images.

Epi-illumination diffraction phase microscopy with white light.

We demonstrate the first reflection-based epi-illumination diffraction phase microscope with white light (epi-wDPM). The epi-wDPM system combines the off-axis, common-path, and white light approaches, in a reflection geometry enabling sub-nanometer spatial and temporal noise levels, while providing single-shot acquisition for opaque samples. We verified the epi-wDPM results by measuring control samples with known dimensions and comparing them to measurements from other well-established techniques. We imaged gold-coated HeLa cells to illustrate the tradeoffs between epi-wDPM with low and high spatial coherence.

Effects of linseeds on the symptoms of irritable bowel syndrome: a pilot randomised controlled trial.

BACKGROUND: Manipulation of dietary fibre intake represents a longstanding treatment for patients with irritable bowel syndrome (IBS), particularly for those with constipation. Linseeds are often recommended by both clinicians and dietitians as a source of dietary fibre to alleviate symptoms. Recent guidance on the management of irritable bowel syndrome (IBS) advises that linseeds may reduce wind and bloating, although there is limited clinical evidence to support this recommendation. The present pilot study aimed to compare the clinical effectiveness of: (i) whole linseeds versus ground linseeds; (ii) whole linseeds versus no linseeds; and (iii) ground linseeds versus no linseeds in the management of IBS symptoms. METHODS: In an open randomised controlled trial, subjects with IBS (n = 40) were allocated to one of three intervention groups: two tablespoons of whole linseeds per day (n = 14), two tablespoons of ground linseeds per day (n = 13) and no linseeds as controls (n = 13). Symptom severity (primary outcome) and bowel habit were assessed before and after a 4-week intervention and statistical differences between the groups were compared. RESULTS: Thirty-one subjects completed the present study. Between-group analysis comparing the improvement in symptom severity did not reach statistical significance for whole linseeds (n = 11) versus ground linseeds (n = 11; P = 0.62), whole linseeds versus controls (n = 9; P = 0.12) and ground linseeds versus controls (P = 0.10). There were no significant changes in stool frequency or stool consistency for any of the groups. CONCLUSIONS: Linseeds may be useful in relief of IBS symptoms. Further research is needed to detect clear differences between the effects of whole and ground linseeds.

British Dietetic Association evidence-based guidelines for the dietary management of irritable bowel syndrome in adults.

BACKGROUND: Irritable bowel syndrome (IBS) is a chronic debilitating functional gastrointestinal disorder. Diet and lifestyle changes are important management strategies. The aim of these guidelines is to systematically review key aspects of the dietary management of IBS, with the aim of providing evidence-based guidelines for use by registered dietitians. METHODS: Questions relating to diet and IBS symptom management were developed by a guideline development group. These included the role of milk and lactose, nonstarch polysaccharides (NSP), fermentable carbohydrates in abdominal bloating, probiotics and empirical or elimination diets. A comprehensive literature search was conducted and relevant studies from January 1985 to November 2009 were identified using the electronic database search engines: Cinahl, Cochrane Library, Embase, Medline, Scopus and Web of Science. Evidence statements, recommendations, good practice points and research recommendations were developed. RESULTS: Thirty studies were critically appraised. A dietetic care pathway was produced following a logical sequence of treatment and formed the basis of these guidelines. Three lines of dietary management were identified. first line: Clinical and dietary assessment, healthy eating and lifestyle management with some general advice on lactose and NSP. Second line: Advanced dietary interventions to improve symptoms based on NSP, fermentable carbohydrates and probiotics. Third line: Elimination and empirical diets. Research recommendations were also identified relating to the need for adequately powered and well designed randomised controlled trials. CONCLUSIONS: These guidelines provide evidence-based details of how to achieve the successful dietary management of IBS.

Safety and cost savings of an improved three-day rule for stool culture in hospitalised children and adults.

Stools sent for culture from patients after three days of hospitalisation have a low yield (<1%) for bacterial enteric pathogens (BEP), excluding Clostridium difficile, and are expensive to process. A 'three-day rule' for rejection of specimens has previously been validated in adults. We evaluated a three-day rule for paediatric stool samples by retrospective review of all stool culture results from 1995 to 2002. Excluding C. difficile, yield for BEP in samples sent within three days following admission was 97/3751 (2.59%) compared with 3/1511 (0.2%) in samples sent more than three days after admission. The criteria for culture would have been met if the rule had been applied for these three samples. We prospectively evaluated potential savings if the rule were applied for both children and adults over a two-month period in 2000. Savings were greater for adults than for children. Of 490 stools from children, 38 (7.8%) samples did not meet the criteria for culture and of 206 stools from adult patients, 64 (31%) did not meet the criteria for culture. We implemented the rule between 1 March 2003 and 31 March 2006. A total of 14 439 stool samples were received from inpatients requesting culture for BEP, excluding C. difficile. Of these, 5744 (39.8%) were rejected because the criteria for culture were not met. This was estimated as an annual saving of 11,848 pounds to the Trust laboratory. If extrapolated to all NHS Trusts, the potential savings could be in the order of 1.18 million pounds annually.

Label-free cell-substrate adhesion imaging on plasmonic nanocup arrays.

Cell adhesion is a crucial biological and biomedical parameter defining cell differentiation, cell migration, cell survival, and state of disease. Because of its importance in cellular function, several tools have been developed in order to monitor cell adhesion in response to various biochemical and mechanical cues. However, there remains a need to monitor cell adhesion and cell-substrate separation with a method that allows real-time measurements on accessible equipment. In this article, we present a method to monitor cell-substrate separation at the single cell level using a plasmonic extraordinary optical transmission substrate, which has a high sensitivity to refractive index changes at the metal-dielectric interface. We show how refractive index changes can be detected using intensity peaks in color channel histograms from RGB images taken of the device surface with a brightfield microscope. This allows mapping of the nonuniform refractive index pattern of a single cell cultured on the plasmonic substrate and therefore high-throughput detection of cell-substrate adhesion with observations in real time.

Insulin-like growth factor-I receptor signalling and acquired resistance to gefitinib (ZD1839; Iressa) in human breast and prostate cancer cells.

De novo and acquired resistance to the anti-tumour drug gefitinib (ZD1839; Iressa), a specific epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) has been reported. We have determined whether signalling through the IGF-I receptor (IGF-1R) pathway plays a role in the gefitinib-acquired resistance phenotype. Continuous exposure of EGFR-positive MCF-7-derived tamoxifen resistant breast cancer cells (TAM-R) to 1 microM gefitinib resulted in a sustained growth inhibition (90%) for 4 months before the surviving cells resumed proliferation. A stable gefitinib-resistant subline (TAM/TKI-R) was established after a further 2 months and this showed no detectable basal phosphorylated EGFR activity. Compared with the parental TAM-R cells, the TAM/ TKI-R cells demonstrated (a) elevated levels of activated IGF-1R, AKT and protein kinase C (PKC)delta, (b) an increased sensitivity to growth inhibition by the IGF-1R TKI AG1024 and (c) an increased migratory capacity that was reduced by AG1024 treatment. Similarly, the EGFR-positive androgen-independent human prostate cancer cell line DU145 was also continuously challenged with 1 microM gefitinib and, although substantial growth inhibition (60%) was seen initially, a gefitinib-resistant variant (DU145/TKI-R) developed after 3 months. Like their breast cancer counterparts, the DU145/TKI-R cells showed increases in the levels of components of the IGF-1R signalling pathway and an elevated sensitivity to growth inhibition by AG1024 compared with the parent DU145 cell line. Additionally, DU145/TKI-R cell migration was also decreased by this inhibitor. We have therefore concluded that in breast and prostate cancer cells acquired resistance to gefitinib is associated with increased signalling via the IGF-1R pathway, which also plays a role in the invasive capacity of the gefitinib-resistant phenotype.

Epidermal growth factor receptor expression by northern analysis and immunohistochemistry in benign and malignant prostatic tumours.

Epidermal growth factor receptor (EGFR) expression in 44 benign prostatic hyperplasia (BPH) and 55 prostatic carcinoma specimens has been investigated using Northern blot analysis and immunohistochemistry. The values obtained for the EGFR mRNA in the BPH and carcinoma specimens were not significantly different and in the latter there was no correlation with grade. In the immunohistochemical assays, two antibodies to the external and one to the internal domain of EGFR were used. The former ones stained the basal cell membranes intensely whilst cytoplasmic staining of secretory epithelium was seen in BPH specimens with the latter. In the carcinoma specimens, the intensity of membrane staining correlated with the two external domain antibodies, r = 0.640, P < 0.001, but neither of these correlated with the EGFR mRNA results. All three antibodies demonstrated a trend towards elevated expression of EGFR with dedifferentiation which reached significance only with the internal domain antibody results, P < 0.02. No correlation was observed with tumour EGFR mRNA values and the EGFR immunohistochemical results. The EGFR immunoreaction with the external domain antibody in 14 treated high-grade tumours was comparable to that obtained in 15 untreated anaplastic prostatic tumours. In 5 patients, both pre- and post-treatment samples were available and these exhibited little or no difference in EGFR expression with therapy.

Evaluation of renal function in potential living kidney donors.

BACKGROUND: Most transplant centers in the United States, including our own, use creatinine clearance (Ccr) to estimate glomerular filtration rate (GFR) in potential living kidney donors. The purpose of this study was to evaluate our experience with estimation of donor GFR by Ccr and to explore other potential strategies for evaluation of renal function in donors. METHODS: Potential donors performed one to three outpatient urine collections for Ccr measurement. Those with low or inconsistent results underwent measurement of GFR by a radionuclide method not dependent upon urine collection (rGFR). The ability to determine adequacy of urine volume by creatinine excretion rate (UVcr) was examined. GFR was also predicted from serum creatinine (Scr) by using two prediction equations [Cockcroft-Gault and that from the Modification of Diet in Renal Disease Study (MDRD)]. The effect of indexing GFR by height, as opposed to body surface area (BSA) was explored. RESULTS: Over a 5-year period, 22 potential donors (approximately 10% of total evaluated) had inconsistent or low Ccr. Most had normal rGFR and, presumably, undercollected their urine. However, several female donors had truly low GFR by rGFR. The accuracy of Ccr (compared with rGFR) was not predicted by UVcr in a clinically useful way. The prediction equations did not have sufficient accuracy or precision to replace rGFR in this population. Indexing GFR estimates by BSA introduced a bias toward lower values in females; this was largely overcome by indexing GFR by height. CONCLUSIONS: Evaluation of GFR in potential living donors by Ccr is not very useful. A more promising strategy could be developed predicting GFR from Scr and indexing the results by height rather than BSA. The optimum method for evaluation of renal function in kidney donors deserves further study.

Comparative analysis of mRNA and protein expression for epidermal growth factor receptor and ligands relative to the proliferative index in human prostate tissue.

The expression of epidermal growth factor receptor (EGF-R), transforming growth factor alpha (TGFalpha), and epidermal growth factor (EGF) was evaluated in a series of prostate cancer (CaP; n = 55) and benign prostate hyperplasia (BPH; n = 44) specimens using immunocytochemistry (ICC) and Northern blotting. In situ hybridization (ISH), performed on a subgroup of these specimens, proved to be a more informative technique for the assessment of messenger RNA (mRNA) in this heterogeneous tissue. A comparative analysis was made in relation to the proliferative index, assessed using the MIB-1 antibody. Elevated levels of EGF-R and TGFalpha, mRNA, and protein were observed in carcinoma cells compared with benign, secretory epithelium using in situ hybridization and immunocytochemistry. In carcinoma specimens evidence of an autocrine growth loop is provided by a correlation between EGF-R and TGFalpha, mRNA (P < .0001), and protein expression (P < .01). A trend toward increased expression of EGF-R and TGFalpha protein with dedifferentiation and a similar trend in the growth fraction suggest a role in tumor progression. Although there was a correlation between EGF-R and the proliferative index (P < .01), no relationship was found between this latter parameter and TGFalpha immunoreactivity (P > .05), indicating that this growth factor may be linked with other aspects of malignant activity rather than directly stimulating proliferation.

Role of autobiographical memory in social problem solving and depression.

Depressed patients frequently exhibit deficiencies in social problem solving (SPS). A possible cause of this deficit is an impairment in patients' ability to retrieve specific autobiographical memories. A clinically depressed group and a hospital control group performed the Means-End Problem-Solving (MEPS; J. J. Platt & G. Spivack, 1975a) task, during which they were required to attend to the memories retrieved during solution generation. Memories were categorized according to whether they were specific, categoric, or extended and whether the valence of the memories was positive or negative. Results support the general hypothesis that SPS skill is a function of autobiographical memory retrieval as measured by a cuing task and by the types of memories retrieved during the MEPS. However, the dysfunctional nature of categoric memories in SPS, rather than the importance of specific memories, was highlighted in the depressed group. Valence proved to be an unimportant variable in SPS ability. The cyclical links among autobiographical memory retrieval, SPS skills, and depression are discussed.

Social problem solving and autobiographical memory in non-clinical depression.

OBJECTIVES: We examine the proposal that specific autobiographical memories may play a major role in successful social problem solving (SPS). DESIGN AND METHODS: Depressed and control students report on the types of memories they retrieve while performing an SPS task and a cueing task. RESULTS: The retrieval of specific memories on the cueing task and during SPS was found to be positively related to SPS skill. CONCLUSIONS: Specific memories are important for successful SPS. However, the relationship between a specific memory deficit and poor SPS is mediated by severity of depression.

The effects of specific retrieval instruction on social problem-solving in depression.

OBJECTIVES: We examine the proposal that social problem-solving in depression may be improved with the retrieval of specific autobiographical memories. DESIGN AND METHODS: Social problem-solving was assessed with the Means-End Problem-Solving task (MEPS; Platt & Spivack, 1975a). Depressed and non-depressed participants were required either to retrieve a specific memory prior to generating a MEPS solution (primed condition) or to report on the memories retrieved during MEPS performance after giving their MEPS solution (non-primed condition). Participants also judged whether the memories retrieved had been helpful or unhelpful for the process of solution generation. RESULTS: In both depressed and non-depressed individuals, priming increased specific memory retrieval but did not improve MEPS performance. An interaction between depression and priming revealed that priming increased the retrieval of helpful memories in the depressed sample. CONCLUSIONS: Specificity is not, in itself, a sufficient retrieval aim for successful social problem-solving. However specific memory priming may be beneficial in depression because it facilitates the recognition of memories which are helpful for problem-solving.

Antibody and complement mediated lysis of felid herpesvirus 1 infected cells in vitro.

The lysis of cells infected by felid herpesvirus 1 (FHV) by feline anti-FHV antibody and complement was demonstrated. Lytic activity was sensitive to dilution of both antibody and, especially, complement. It was first detected within 10 to 20 minutes, increased rapidly during the next 30 minutes of incubation and then rose more slowly in a linear manner. Using standard antibody and complement concentrations and assay duration, it was shown that FHV infected cells underwent significant (P less than 0.05) lysis from eight hours after infection in a system in which FHV-specific membrane antigen was first detected at three hours after infection and spread of FHV by the intracellular route began eight to nine hours after infection. The ability of antibody and complement to reduce FHV spread in this system was demonstrated by a significant (P less than 0.05) reduction in FHV plaque numbers, although the restriction of spread was not absolute. Chelation of divalent cations and heat inactivation of complement factor B revealed that the lytic system was dependent on factor B and Mg2+ but not Ca2+, suggesting involvement of the alternative pathway of complement activation.

Gender differences in the dual-task effects on autobiographical memory retrieval during social problem solving.

A dual-task paradigm was used to explore the effects of cognitive load on social problem solving and autobiographical memory retrieval. The role that gender may play in mediating the relationship was also examined. Participants performed a secondary task concurrently with two primary tasks: (a) a cueing task, and (b) the Means-End Problem-Solving (MEPS) Task, during which they were required to attend to the memories retrieved during solution generation. Two dual-task conditions were employed in order that two levels of secondary task difficulty could be explored. Level of difficulty proved to be an important factor in the effects of resource reduction on the two primary tasks. Retrieval during the MEPS was effected by both the easy and difficult secondary task whereas retrieval on the cueing task was affected by the difficult task only. The results also showed that females (in contrast to males) favoured a more detailed SPS style using a specific memory database. Consequently, under central executive pressure, females' performance was significantly affected while males' performance remained largely unchanged.

Sexuality and spinal cord injury.

Spinal cord injury may have a profound effect on the patient's sexuality. In order to maximize the patient's potential, concern and support from health care professionals is needed. This article reviews normal sexual function and changes occurring following spinal cord injury. The PLISSIT model is described and is used to describe nursing interventions designed to deal with the sexual concerns of spinal cord-injured patients.

Changing attitudes about persons with disabilities: effects of a simulation.

The question of whether the use of a teaching strategy employing simulation influences attitudes toward disabled persons over time was addressed in this quasi-experimental intervention study. The experimental group participated in "Sensitivity Day" which incorporated a simulation activity. A total of 77 undergraduate baccalaureate nursing students completed the Attitudes Toward Disabled Persons scale four times: presimulation, immediate postsimulation, six week delayed postsimulation and six month delayed postsimulation. In general, the nursing students had relatively positive scores on the Attitudes Toward Disabled Persons scale in each testing period. The best indicator of postsimulation attitude scores was the presimulation attitude score. Personal and prior nursing experience had no statistical effects on attitude. Improvement that was significant at p = .07 level was noted in the experimental group at the six weeks delayed postsimulation when compared with the presimulation attitude scores. This study supplies data that is useful in constructing learning activities and contributes to the body of knowledge about simulation and attitudes toward persons with disabilities.

A head-injured patient: caloric needs, clinical progress and nursing care priorities.

This case study explored literature on nursing care of the head-injured patient and identified outcomes of a head-injured patient in regard to nutritional status, skin breakdown and infection. Literature review revealed head-injured patients experience a hypermetabolism requiring increased nutritional support. Patient outcomes included mild skin breakdown, infection and a 12% loss in body weight. Inadequate nutrition was the patient's most significant outcome. The patient maintained an average daily intake of 915 calories and averaged a total energy expenditure of 2337 calories, thus, demonstrating an average daily deficit of 1422 calories. Top nursing care priorities noted in the patient's chart were: alteration in cerebral tissue perfusion, potential for injury and alteration in skin integrity. Results indicated more accurate assessment of the patient's needs and inclusion of more applicable nursing diagnoses were required to ensure greater continuity in care.