This study has utilised comparative functional genomics to exploit animal models with extreme adaptation to lactation to identify candidate genes that specifically regulate protein synthesis in the cow mammary gland. Increasing milk protein production is valuable to the dairy industry. The lactation strategies of both the Cape fur seal (Artocephalus pusillus pusillus) and the tammar wallaby (Macropus eugenii) include periods of high rates of milk protein synthesis during an established lactation and therefore offer unique models to target genes that specifically regulate milk protein synthesis. Global changes in mammary gene expression in the Cape fur seal, tammar wallaby, and the cow (Bos taurus) were assessed using microarray analysis. The folate receptor alpha (FOLR1) showed the greatest change in gene expression in all three species [cow 12.7-fold (n = 3), fur seal 15.4-fold (n = 1), tammar 2.4-fold (n = 4)] at periods of increased milk protein production. This compliments previous reports that folate is important for milk protein synthesis and suggests FOLR1 may be a key regulatory point of folate metabolism for milk protein synthesis within mammary epithelial cells (lactocytes). These data may have important implications for the dairy industry to develop strategies to increase milk protein production in cows. This study illustrates the potential of comparative genomics to target genes of interest to the scientific community.

As synthetic biology matures to compete with chemical transformation of commodity and high-value compounds, a wide variety of well-characterized biological parts are needed to facilitate system design. Protein quantification based on selected-reaction monitoring (SRM) mass spectrometry compliments metabolite and transcript analysis for system characterization and optimizing flux through engineered pathways. By using SRM quantification, we assayed red fluorescent protein (RFP) expressed from plasmids containing several inducible and constitutive promoters and subsequently assessed protein production from the same promoters driving expression of eight mevalonate pathway proteins in Escherichia coli. For each of the promoter systems, the protein level for the first gene in the operon followed that of RFP, however, the levels of proteins produced from genes farther from the promoter were much less consistent. Second, we used targeted proteomics to characterize tyrosine biosynthesis pathway proteins after removal of native regulation. The changes were not expected to cause significant impact on protein levels, yet significant variation in protein abundance was observed and tyrosine production for these strains spanned a range from less than 1 mg/L to greater than 250 mg/L. Overall, our results underscore the importance of targeted proteomics for determining accurate protein levels in engineered systems and fine-tuning metabolic pathways.

BACKGROUND

The recent development of novel repeat-fruiting types of blackberry (Rubus L.) cultivars, combined with a long history of morphological marker-assisted selection for thornlessness by blackberry breeders, has given rise to increased interest in using molecular markers to facilitate blackberry breeding. Yet no genetic maps, molecular markers, or even sequences exist specifically for cultivated blackberry. The purpose of this study is to begin development of these tools by generating and annotating the first blackberry expressed sequence tag (EST) library, designing primers from the ESTs to amplify regions containing simple sequence repeats (SSR), and testing the usefulness of a subset of the EST-SSRs with two blackberry cultivars.

RESULTS

A cDNA library of 18,432 clones was generated from expanding leaf tissue of the cultivar Merton Thornless, a progenitor of many thornless commercial cultivars. Among the most abundantly expressed of the 3,000 genes annotated were those involved with energy, cell structure, and defense. From individual sequences containing SSRs, 673 primer pairs were designed. Of a randomly chosen set of 33 primer pairs tested with two blackberry cultivars, 10 detected an average of 1.9 polymorphic PCR products.

CONCLUSIONS

This rate predicts that this library may yield as many as 940 SSR primer pairs detecting 1,786 polymorphisms. This may be sufficient to generate a genetic map that can be used to associate molecular markers with phenotypic traits, making possible molecular marker-assisted breeding to compliment existing morphological marker-assisted breeding in blackberry.

Prenatal exposure to persistent organic pollutants (POPs), may influence offspring weight gain. More prospective epidemiological studies are needed to compliment the growing body of evidence from animal studies.

Serum from 412 pregnant Norwegian and Swedish women participating in a Scandinavian prospective cohort study were collected in 1986-88, and analyses of two perfluoroalkyl substances (PFASs) and five organochlorines (OCs) were conducted. We used linear and logistic regression models with 95% confidence intervals (CIs) to evaluate the associations between maternal serum POP concentrations at 17-20 weeks of gestation and child overweight/obesity (body mass index (BMI) ≥ 85th percentile) at 5-year follow-up. Results were further stratified by country after testing for effect modification. We also assessed potential non-monotonic dose-response (NMDR) relationships.

In adjusted linear models, we observed increased BMI-for-age-and-sex z-score (β = 0.18, 95% CI: 0.01-0.35), and increased triceps skinfold z-score (β = 0.15, 95% CI: 0.02-0.27) in children at 5-year follow-up per ln-unit increase in maternal serum perfluorooctane sulfonate (PFOS) concentrations. We observed increased odds for child overweight/obesity (BMI ≥ 85th percentile) for each ln-unit increase in maternal serum PFOS levels (adjusted OR: 2.04, 95% CI: 1.11-3.74), with stronger odds among Norwegian children (OR: 2.96, 95% CI: 1.42-6.15). We found similar associations between maternal serum perfluorooctanoate (PFOA) concentrations and child overweight/obesity. We found indications of NMDR relationships between PFOS and polychlorinated biphenyl (PCB) 153 and child overweight/obesity among Swedish children.

We found positive associations between maternal serum PFAS concentrations and child overweight/obesity at 5-year follow-up, particularly among Norwegian participants. We observed some evidence for NMDR relationships among Swedish participants.

Centuries since it was first described, tuberculosis (TB) remains a significant global public health issue. Despite ongoing holistic measures implemented by health authorities and a number of new oral treatments reaching the market, there is still a need for an advanced, efficient TB treatment. An adjunctive, host-directed therapy designed to enhance endogenous pathways and hence compliment current regimens could be the answer. The integration of drug repurposing, including synthetic and naturally occurring compounds, with a targeted drug delivery platform is an attractive development option. In order for a new anti-tubercular treatment to be produced in a timely manner, a multidisciplinary approach should be taken from the outset including stakeholders from academia, the pharmaceutical industry, and regulatory bodies keeping the patient as the key focus. Pre-clinical considerations for the development of a targeted host-directed therapy are discussed here.

BACKGROUND

Psychosocial cancer research illustrates how women treated for breast cancer experience physical changes in their bodies and the way they perceive, that, others see their body. But how patients with other types of cancer have experienced changes in their bodies and how this affects their relationship with others is less researched.

OBJECTIVE

To explore how cancer survivors with different types of cancer and cancer treatment, experience and handle their changed body, especially when meeting others, and how this influences their everyday life of survivorship, i.e. long after treatment has been completed.

METHODS

Participant observation at a Cancer Rehabilitation Centre (CRC). Of the observed participants 23 were selected and interviewed twice.

RESULTS

Many participants had a changed body due to the cancer and its treatment. When the cancer survivors meet others they experience that their changed body means that they are avoided, looked at in specific ways, or greeted with a specific compliment. The verbal and nonverbal language that the cancer survivors are met with indicates the existence of a specific discursive aesthetic in relation to the disease and the changed body. This discursive aesthetic represents a silence and secrecy about cancer, which makes it impossible for survivors to talk about their experiences with cancer and a changed body.

CONCLUSIONS

The changed body not only represents the physical sign of cancer, it also represents the social presence and representation of cancer. The analysis gives an insight into general questions of meaning related to the changed body in late modernity.

Across the rostrocaudal (RC) axis of the Xenopus tadpole optic tectum exists a developmental gradient. This gradient has served as a useful model to study many aspects of synapse and dendrite maturation. To compliment these studies, we characterized how the intrinsic excitability, the ease in which a neuron can fire action potentials, might also be changing across the same axis. Whole-cell recordings from tectal neurons at different points along the RC axis revealed a graded increase in intrinsic excitability: compared with neurons at the caudal end of the tectum, neurons at the rostral end fired more action potentials in response to current injection and expressed greater peak Na⁺ and K⁺ currents, the major intrinsic currents in these neurons that underlie the action potential. We also observed, along the same axis and in the same direction, a previously described increase in the amount of synaptic drive received by individual neurons (Wu GY, Malinow R, Cline HT. Science 274: 972-976, 1996). Thus as synaptic activity ramps up across the RC axis, so does intrinsic excitability. The reduction of overall circuit activity induced a compensatory scaling up of peak Na⁺ and K⁺ currents only in the caudal portion of the tectum, suggesting a region-specific, compensatory form of plasticity.

The coronary artery bypass graft (CABG) operation is one of the few remaining operations/interventions on diseased arteries that are not routinely verified during or immediately after the procedure. This review answers the 'how', 'when' and 'why' of intraoperative CABG assessment.

More recent than new literature on this topic, is the increased interest in quality assurance of CABG. This is most likely due to reports in the last 5 years suggesting CABG superiority to percutaneous coronary intervention (PCI) for improved mid-term and long-term outcomes; for example, for patients with diabetes mellitus (Freedom Trial by Farkouh in 2012), and for patients with SYNTAX score ≥ 33 (SYNTAX Trial by Mohr in 2013). Possibly CABG is re-emerging from the era-of-better-and-better-stents and is now deemed worthy of improvement.

In order to fully compliment PCI, the operative major adverse cardiac event rate of CABG must rival that of PCI. In order to reduce technical errors, it is best practice to perform intra-operative assessment of bypasses, especially since we have the tools.

In the last 25 yr, there have been several advances in the safe management of the airway. Videolaryngoscopes and supraglottic airways, now in routine use by new trainees in anaesthesia, have had their genesis in the recent past. The 4th National Audit Project of the Royal College of Anaesthetists and the Difficult Airway Society published in 2011 a seminal report that has influenced airway management worldwide . Understanding how the report's recommendations were constructed and how clinical guidelines compliment rather than contradict them is important in understanding the tenets of safe airway management. Over the last 25 yr there has been an increasing understanding of the effects of human factors in anaesthesiology: we may not perform in a predictable or optimal manner when faced with unusual and threatening challenges. The place of cricoid pressure in anaesthetic practice has also evolved. Current recommendations are that it be applied, but it should be released rapidly should airway difficulty be encountered. The need to prevent hypoxaemia by preoxygenation has long been recognized, but the role of high-flow nasal oxygen in anaesthesia is now being realized and developed. Clinicians must decide how novel therapies and long-standing practices are adapted to best meet the needs of our patients and prevent harm during airway management.

OBJECTIVE

High-resolution pharyngeal manometry (HRPM) is a well tolerated, catheter-based, method for recording the pressures and bolus flow generated by the pharyngeal muscles during swallowing. Despite a body of published evidence and a critical mass of investigators in the field, there is a lack of consensus surrounding what biomechanical phenomena to measure. The purpose of this review is to provide some insights into the information on swallowing physiology that can be gathered using HRPM.

RESULTS

HRPM literature has focused on measuring biomechanical phenomena that may be relevant to measure in relation to dysphagia research. This review focuses on the measurement of pharyngeal luminal occlusive forces, intrabolus distension pressure, bolus presence and bolus flow timing as key features of pharyngeal swallowing that require measurement and allow for derivation of the Swallow Risk Index, a global measure of swallow function indicative of swallowing functional reserve.

CONCLUSIONS

HRPM allows objective derivation of measures of swallow function that may have value for diagnosis and research in relation to swallowing disorders. HRPM has demonstrated clinical applicability in specific patient populations and offers unique advantages that compliment current assessment methods.

Understanding the pathophysiology and pharmacology of migraine has been driven by astute clinical observations, elegant experimental medicine studies, and importantly by studying highly effective anti-migraine agents in the laboratory and the clinic. Significant progress has been made in the use of functional brain imaging to compliment observational studies of migraine phenotypes by highlighting pathways within the brain that may be involved in predisposition to migraine, modulating migraine pain or that could be sensitive to pharmacological or behavioral therapeutic intervention. In drug discovery, molecular imaging approaches compliment functional neuroimaging by visualizing migraine drug targets within the brain. Molecular imaging enables the selection and evaluation of drug candidates by confirming that they engage their targets sufficiently at well tolerated doses to test our therapeutic hypotheses. Migraine is a progressive disorder. Developing our knowledge of where drugs act in the brain and of how the brain is altered in both episodic migraine (interictal state and ictal state) and chronic migraine are important steps to understanding why there is such differential responsiveness to therapeutics among migraine patients and to improving how they are evaluated and treated.

OBJECTIVE

To evaluate whether a mobile health application that employs elements of social game design could compliment medical care for unresolved concussion symptoms.

METHODS

Phase I and Phase II (open-label, non-randomized, ecological momentary assessment methodology).

METHODS

Outpatient concussion clinic.

METHODS

Youth, aged 13-18 years, with concussion symptoms 3+ weeks after injury; Phase I: n = 20; Phase II: n = 19.

METHODS

Participants received standard of care for concussion. The experimental group also used a mobile health application as a gamified symptoms journal.

METHODS

Phase I: feasibility and satisfaction with intervention (7-point Likert scale, 1 high). Phase II: change in SCAT-3 concussion symptoms (primary), depression and optimism.

RESULTS

Phase 1: A plurality of participants completed the intervention (14 of 20) with high use (110 +/- 18% play) and satisfaction (median +/- interquartile range (IQR) = 2.0+/- 0.0). Phase II: Groups were equivalent on baseline symptoms, intervention duration, gender distribution, days since injury and medication prescription. Symptoms and optimism improved more for the experimental than for the active control cohort (U = 18.5, p = 0.028, effect size r = 0.50 and U = 18.5, p = 0.028, effect size r = 0.51, respectively).

CONCLUSIONS

Mobile apps incorporating social game mechanics and a heroic narrative may promote health management among teenagers with unresolved concussion symptoms.

This study traces the biosocial dynamics of Echinococcus granulosus - a zoonotic tapeworm spread between dogs, livestock and people - at slaughterhouses in Morocco. One of the most important parasitic zoonoses worldwide, this neglected cestode is responsible for a debilitating, potentially life-threatening, human disease and significant livestock production losses. Transmission can be interrupted, among other ways, by restricting dogs from eating cyst-infected livestock viscera. Recent epidemiological studies in Sidi Kacem province, northern Morocco, found that government-operated slaughterhouses were 'hotspots' for hydatid cysts in livestock and infection in dogs. An ethnographic approach was used to compliment these studies, exploring 'how' and 'why' cysts were being openly discarded. All seven visited slaughterhouses had low levels of hygiene, oversight and infrastructure. This was described locally as perpetuating a sense of 'chaos' that normalized (un)hygienic practices and justified the ignoring of state rules and regulations. However deference to 'poor' infrastructure, both physically and symbolically, served to under-emphasize local institutional logics, which were mediated by prevailing risk perceptions, economic practices and local socio-political norms. These included inter-departmental government relationships, the motivation of veterinary technicians, the political lobbying of butchers and market-based mitigation strategies. The study shows the importance of understanding E. granulosus from a biosocial perspective, and the need for more long-term, participatory and integrated 'One Health' approaches for neglected zoonotic diseases.

OBJECTIVE

Modern neonatal care, surgical treatment, and total parenteral nutrition (TPN) have improved survival rate for babies with extensive gut resections. The authors examined the role of intestinal transplantation in the treatment of these patients.

METHODS

The authors reviewed all pediatric intestinal transplants performed for short bowel syndrome at our center (70 transplants performed between Aug 1994 and Feb 2002). Factors affecting patient survival were analyzed.

RESULTS

Older patient age at the time of transplant was a significant factor favorably affecting patient survival (P =.031). Trends toward better survival rates were observed in those transplants performed more recently (P =.063), in those patients with greater body weight (P =.084), in those not hospitalized at the time of transplant (P =.14), and in those without concomitant liver failure (P =.12). Three-year survival rate for patients greater than age 2 years and without liver failure was 90%. However, 32% of our recipients underwent transplant at age less than one year, and most in this group (75%) had concomitant liver failure.

CONCLUSIONS

For babies with irreversible intestinal failure, intestinal transplantation is a life-saving option. Results, which have recently improved, are best when transplantation compliments more conservative surgical treatments and TPN. However, there is a subset of patients who have liver disease early requiring urgent transplant.

Roseobacter is a dominant lineage in the marine environment. This group of bacteria is diverse in terms of both their phylogenetic composition and their physiological potential. Roseobacter denitrificans OCh114 is one of the most studied bacteria of the Roseobacter lineage. Recently, a lytic phage (RDJLPhi1) that infects this bacterium was isolated and a mutant strain (M1) of OCh114 that is resistant to RDJLPhi1 was also obtained. Here, we investigate the mechanisms supporting phage resistance of M1. Our results excluded the possibilities of several phage resistance mechanisms, including abortive infection, lysogeny, and the clustered regularly interspaced short palindromic repeats (CRISPRs) related mechanism. Adsorption kinetics assays revealed that adsorption inhibition might be a potential cause for the phage resistance of M1. Comparative proteomic analysis of M1 and OCh114 revealed significant changes in the membrane protein compliment of these bacteria. Five membrane proteins with important biological functions were significantly down-regulated in the phage-resistant M1. Meanwhile, several outer membrane porins with different modifications and an OmpA family domain protein were markedly up-regulated. We hypothesize that the down-regulated membrane proteins in M1 may serve as the potential phage receptors, whose absence prevented the adsorption of phage RDJLPhi1 to host cells and subsequent infection.

We investigated children's understanding of irony and sensitivity to irony's meanness and humor. In Study 1, 89 participants (5-6-year-olds, 8-9-year-olds, adults) heard ironic and literal criticisms, and literal compliments. Comprehension of irony emerged between 5 and 6 years of age. Ratings of humor increased with age; ratings of meanness did not (showing that all ages perceived irony as more muted than literal criticism). In Study 2, results from 135 participants (6-7-year-olds, 8-9-year-olds, and adults) replicated these findings and revealed the role of form and intonation. Thus, comprehension of irony emerges between 5 and 6 years of age, and sensitivity to the muting function develops prior to sensitivity to the humor function.

Despite use of meticulous surgical techniques, and regardless of surgical access via laparotomy or laparoscopy, postoperative adhesions develop in the vast majority of women undergoing abdominopelvic surgery. Such adhesions represent not only adhesion reformation at sites of adhesiolysis, but also de novo adhesion formation at sites of surgical procedures. Application of antiadhesion adjuvants compliment the benefits of meticulous surgical techniques, providing an opportunity to further reduce postoperative adhesion development. Improved understanding of the pathophysiology of adhesion development and distinguishing variations in the molecular biologic mechanisms from adhesion-free peritoneal repair represent future opportunities to improve the reduction of postoperative adhesions. Optimization of the reduction of postoperative adhesions will likely require identification of unique, personalized approaches in each individual, representing interindividual variation in peritoneal repair processes.

BACKGROUND

The 2 Cohort randomised PrefHer trial examined the preferences of HER2+ve primary breast cancer patients for intravenous (IV) or subcutaneous (SC) delivery of trastuzumab via a Single Injectable Device (SID) or hand-held syringe (HHS). The novel approach and design of the study permitted an in-depth exploration of patients' experiences, the impact that different modes of delivery had on patients' well-being and implications for future management.

METHODS

The preferences, experiences and general comments of patients in the PrefHer study were collected via specific semi-structured interview schedules. Exploratory analyses of data were conducted using standard methodology. The final question invited patients to make further comments, which were divided into 9 thematic categories - future delivery, compliments, time/convenience, practical considerations, pain/discomfort, study design, side-effects, psychological impact, and perceived efficacy.

RESULTS

267/467 (57%) patients made 396 additional comments, 7 were neutral, 305 positive and 86 negative. The three top categories generating the largest number of comments were compliments and gratitude about staff and being part of PrefHer (75/396; 19%), the potential future delivery of SC trastuzumab (73/396; 18%), and practical considerations about SC administration (60/396; 15%).

CONCLUSIONS

Eliciting patient preferences about routes of administration of drugs via comprehensive interviews within a randomised cross-over trial yielded rich and important information. The few negative comments made demonstrated a need for proper staff training in SC administration Patients were grateful to have been part of the trial, and would have liked to continue with SC delivery. The possibility of home administration in the future also seemed acceptable.

UNASSIGNED

2010-024099-25.

BACKGROUND

An essential developmental pathway in sexually reproducing animals is the specification of germ cells and the differentiation of mature gametes, sperm and oocytes. The "germline" genes vasa, nanos and piwi are commonly identified in primordial germ cells, suggesting a molecular signature for the germline throughout animals. However, these genes are also expressed in a diverse set of somatic stem cells throughout the animal kingdom leaving open significant questions for whether they are required for germline specification. Similarly, members of the Dmrt gene family are essential components regulating sex determination and differentiation in bilaterian animals, but the functions of these transcription factors, including potential roles in sex determination, in early diverging animals remain unknown. The phylogenetic position of ctenophores and the genome sequence of the lobate Mnemiopsis leidyi motivated us to determine the compliment of these gene families in this species and determine expression patterns during development.

RESULTS

Our phylogenetic analyses of the vasa, piwi and nanos gene families show that Mnemiopsis has multiple genes in each family with multiple lineage-specific paralogs. Expression domains of Mnemiopsis nanos, vasa and piwi, during embryogenesis from fertilization to the cydippid stage, were diverse, with little overlapping expression and no or little expression in what we think are the germ cells or gametogenic regions. piwi paralogs in Mnemiopsis had distinct expression domains in the ectoderm during development. We observed overlapping expression domains in the apical organ and tentacle apparatus of the cydippid for a subset of "germline genes," which are areas of high cell proliferation, suggesting that these genes are involved with "stem cell" specification and maintenance. Similarly, the five Dmrt genes show diverse non-overlapping expression domains, with no clear evidence for expression in future gametogenic regions of the adult. We also report on splice variants for two Mnemiopsis Dmrt genes that impact the presence and composition of the DM DNA binding domain for these transcription factors.

CONCLUSIONS

Our results are consistent with a potential role for vasa, piwi and nanos genes in the specification or maintenance of somatic stem cell populations during development in Mnemiopsis. These results are similar to previous results in the tentaculate ctenophore Pleurobrachia, with the exception that these genes were also expressed in gonads and developing gametes of adult Pleurobrachia. These differences suggest that the Mnemiopsis germline is either specified later in development than hypothesized, the germline undergoes extensive migration, or the germline does not express these classic molecular markers. Our results highlight the utility of comparing expression of orthologous genes across multiple species. We provide the first description of Dmrt expression in a ctenophore, which indicates that Dmrt genes are expressed in distinct structures and regions during development but not in future gametogenic regions, the only sex-specific structure for this hermaphroditic species.

This paper reviews procedures for ensuring that physicians in the European Union (EU) continue to meet criteria for registration and the implications of these procedures for cross-border movement of health professionals following implementation of the 2005/36/EC Directive on professional qualifications. A questionnaire was completed by key informants in 10 EU member states, supplemented by a review of peer-reviewed and grey literature and a review conducted by key experts in each country. The questionnaire covered three aspects: actors involved in processes for ensuring continued adherence to standards for registration and/or licencing (such as revalidation), including their roles and functions; the processes involved, including continuing professional development (CPD) and/or continuing medical education (CME); and contextual factors, particularly those impacting professional mobility. All countries included in the study view CPD/CME as one mechanism to demonstrate that doctors continue to meet key standards. Although regulatory bodies in a few countries have established explicit systems of ensuring continued competence, at least for some doctors (in Belgium, Germany, Hungary, the Netherlands, Slovenia and the UK), self-regulation is considered sufficient to ensure that physicians are up to date and fit to practice in others (Austria, Finland, Estonia and Spain). Formal systems vary greatly in their rationale, structure, and coverage. Whereas in Germany, Hungary and Slovenia, systems are exclusively focused on CPD/CME, the Netherlands also includes peer review and minimum activity thresholds. Belgium and the UK have developed more complex mechanisms, comprising a review of complaints or compliments on performance and (in the UK) colleague and patient questionnaires. Systems for ensuring that doctors continue to meet criteria for registration and licencing across the EU are complex and inconsistent. Participation in CPD/CME is only one aspect of maintaining professional competence but it is the only one common to all countries. Thus, there is a need to bring clarity to this confused landscape.

Local treatment of lung cancer using inhalable nanoparticles (NPs) is an emerging and promising treatment option. The aim of this study was to investigate the activation of alveolar macrophages by poly (isobutyl cyanoacrylate) (BIPCA) NPs and the consequences of this activation on H460 lung cancer cells. A methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay was used to determine the primary cytotoxicity, that is, the immediate and direct cytotoxicity of doxorubicin (DOX)-loaded NPs on both cell lines. Macrophages were then treated using EC(50) concentrations of different treatments and co-cultured in a two-compartment system with H460 lung cancer cells. These treatments included DOX solution, blank NPs, and DOX-loaded NPs. The results showed that alveolar macrophages exposed to blank or DOX-loaded NPs showed cytotoxicity against cancer cells after 8 and 24h; this behavior was not expressed by naïve macrophages or macrophages treated with DOX solution. Sample analysis indicated that macrophages have the ability to release back fragments of NPs that were previously phagocytized. Further investigations showed that NPs can induce an increase in the excretion of Th1 cytokines namely, monocytes chemoattractant protein-1 (MCP-1), macrophages inflammatory protein (MIP-1), tumor necrosis factor alpha (TNF-alpha), and interferon gamma (IFN-gamma). The Th1 cytokines released by the alveolar macrophages might explain the significant secondary cytotoxicity effect on H460 cancer cells. Secondary cytotoxicity mediated by macrophages might compliment the direct cytotoxic effect that NPs have on cancer cells.

DNA microarrays can give global transcriptional views of cellular responses to disease, development, nutrition, and other biological states. They can be used to elucidate biological networks, develop diagnostics, and identify genetic targets and molecular mechanisms. The technology is widely used and can be a valuable complement to more "disease-centric" focused arrays. For these reasons, Nestlé designed a custom canine Affymetrix microarray representing transcripts from multiple tissues for use in areas where a more focused microarray had not already been developed. Sufficient numbers of sequences representing messenger RNAs (mRNAs) or expressed sequence tags (ESTs) is integral for the design of a global microarray chip. This chip was designed using public domain sequences (GenBank) and sequences from a proprietary canine EST database. In order to enrich the chip with annotated transcripts, both of these sequence sets were BLASTed against the nonredundant protein database. The sequences on the microarray were isolated from more than 48 different tissues. The final compliment of sequences had sequences unique to GenBank (3160), unique to the proprietary EST database (17,620), and present in both sources (1996). In comparison with human sequences (RefSeq), 74% of the canine sequences matched a human sequence.

OBJECTIVE

reduced fetal movements (RFM) are experienced by 46-50% of women prior to the diagnosis of stillbirth. Empowering women with evidenced-based information regarding RFM may allow for prompt contact with a health care provider and access to appropriate management. Use of the Internet is growing in popularity as a source of pregnancy information to aid mothers׳ decision-making. This study aimed to identify and examine the available online information for pregnant women regarding RFM.

METHODS

a systematic search was performed using Google, Yahoo and Bing to identify the most popular websites giving information about RFM. The websites were assessed for readability, accountability and content using the Flesh-Kincaid ease of readability score; the Silberg criteria; and by comparison to evidence-based guidelines respectively. Chat forums were assessed using a qualitative thematic analysis.

RESULTS

70 information articles and 63 chat forums were analysed from 77 unique websites. The mean readability score was 65.7 (suitable for the average 13-15 year old) and therefore above the recommended level set for health materials; only 15 (21.4%) websites met all accountability criteria; and 43 (70%) websites contained information that was not in accordance with evidence-based recommendations. Typical questions on forums were 'Is this normal? What should I do?' and responses were 'Better safe than sorry', 'There is no harm in calling'.

CONCLUSIONS

overall, there was wide variation in the quality of information regarding RFM on the Internet. However, the study identified four excellent websites on RFM that may be suitable sources of information for women. Women׳s uncertainty displayed in the chat forums may suggest that clearer, accessible guidance is needed if they experience RFM.

CONCLUSIONS

the Internet can compliment and support current methods of antenatal information provision. However, due to varying levels of quality it is essential that professionals discuss and direct women to useful evidenced-based websites. Care must be taken to ensure consistent advice and management of those presenting with RFM and that women׳s concerns are addressed.

Surface-enhanced laser desorption/ionization (SELDI) time-of-flight (TOF) mass spectrometry (MS) has been widely applied for conducting biomarker research with the goal of discovering patterns of proteins and/or peptides from biological samples that reflect disease status. Many diseases, ranging from cancers of the colon, breast, and prostate to Alzheimer's disease, have been studied through serum protein profiling using SELDI-based methods. Although the results from SELDI-based diagnostic studies have generated a great deal of excitement and skepticism alike, the basis of the molecular identities of the features that underpin the diagnostic potential of the mass spectra is still largely unexplored. A detailed investigation has been undertaken to identify the compliment of serum proteins that bind to the commonly used weak cation exchange (WCX-2) SELDI protein chip. Following incubation and washing of a standard serum sample on the WCX-2 sorbent, proteins were harvested, digested with trypsin, fractionated by strong cation exchange liquid chromatography (LC), and subsequently analyzed by microcapillary reversed-phase LC coupled online with an ion-trap mass spectrometer. This analysis resulted in the identification of 383 unique proteins in the WCX-2 serum retentate. Among the proteins identified, 50 (13%) are documented clinical biomarkers with 36 of these (72%) identified from multiple peptides.