Isolation and Profiling of Circulating Tumorâ Associated Exosomes Using Extracellular Vesicular Lipidâ Protein Binding Affinity Based Microfluidic Device

Extracellular vesicles (EVs) are emerging as a potential diagnostic test for cancer. Owing to the recent advances in microfluidics, onâ chip EV isolation is showing promise with respect to improved recovery rates, smaller necessary sample volumes, and shorter processing times than ultracentrifugation. Immunoaffinityâ based microfluidic EV isolation using antiâ CD63 is widely used; however, antiâ CD63 is not specific to cancerâ EVs, and some cancers secrete EVs with low expression of CD63. Alternatively, phosphatidylserine (PS), usually expressed in the inner leaflet of the lipid bilayer of the cells, is shown to be expressed on the outer surface of cancerâ associated EVs. A new exosome isolation microfluidic device (newExoChip), conjugated with a PSâ specific protein, to isolate cancerâ associated exosomes from plasma, is presented. The device achieves 90% capture efficiency for cancer cell exosomes compared to 38% for healthy exosomes and isolates 35% more A549â derived exosomes than an antiâ CD63â conjugated device. Immobilized exosomes are then easily released using Ca2+ chelation. The recovered exosomes from clinical samples are characterized by electron microscopy and westernâ blot analysis, revealing exosomal shapes and exosomal protein expressions. The newExoChip facilitates the isolation of a specific subset of exosomes, allowing the exploration of the undiscovered roles of exosomes in cancer progression and metastasis.Onâ chip exosome isolation using immunoaffinity capture improves recovery from small sample volumes compared to ultracentrifugation. However, tetraspanins are not specific to cancer exosomes and some cancers secrete exosomes with low expression of tetraspanins. Here, a microfluidicâ device conjugated with proteins against phosphatidylserine (PS), shown to be expressed on the outer surface of cancerâ associated exosome, is used to isolate cancerâ associated exosomes.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/152534/1/smll201903600-sup-0001-SuppMat.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/152534/2/smll201903600_am.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/152534/3/smll201903600.pd

A Systematic Review of Behavioral Interventions for Elementary School Children with Social, Emotional and Behavioral Difficulties: Contributions from Single-Case Research Studies

Challenges arising from the classroom behavioral management of students with social, emotional and behavioral difficulties are a concern for educational professionals. The purpose of this study is to review common elements of behavior interventions for the disruptive behaviors of children with social, emotional, and behavioral difficulties. A systematic review was conducted through an electronic search of studies (from 2000 to 2017) on ERIC, Web of Science, FRANCIS, and MEDLINE databases. The inclusion criteria involved: (i) an intervention improving behaviors at school of children with disruptive behaviors; (ii) elementary school children with the majority of the sample or average age between 6-11 years old; (iii) at least one measurable outcome focusing on social/emotional/behavioral outcomes; (iv) single-case designs. Of the 5339 articles that were identified in the initial screening, 27 met the criteria to be included in the review. Common characteristics of successful interventions are discussed to make recommendations for future implementation.info:eu-repo/semantics/publishedVersio

Self-report underestimates the frequency of the acute respiratory exacerbations of COPD but is associated with BAL neutrophilia and lymphocytosis: an observational study.

RATIONALE: Research studies typically quantify acute respiratory exacerbation episodes (AECOPD) among people with chronic obstructive pulmonary disease (COPD) based on self-report elicited by survey questionnaire. However, AECOPD quantification by self-report could be inaccurate, potentially rendering it an imprecise tool for identification of those with exacerbation tendency. OBJECTIVE: Determine the agreement between self-reported and health records-documented quantification of AECOPD and their association with airway inflammation. METHODS: We administered a questionnaire to elicit the incidence and severity of respiratory exacerbations in the three years preceding the survey among current or former heavy smokers with or without diagnosis of COPD. We then examined electronic health records (EHR) of those with COPD and those without (tobacco-exposed persons with preserved spirometry or TEPS) to determine whether the documentation of the three-year incidence of moderate to very severe respiratory exacerbations was consistent with self-report using Kappa Interrater statistic. A subgroup of participants also underwent bronchoalveolar lavage (BAL) to quantify their airway inflammatory cells. We further used multivariable regressions analysis to estimate the association between respiratory exacerbations and BAL inflammatory cell composition with adjustment for covariates including age, sex, height, weight, smoking status (current versus former) and burden (pack-years). RESULTS: Overall, a total of 511 participants completed the questionnaire, from whom 487 had EHR available for review. Among the 222 participants with COPD (70 ± 7 years-old; 96% male; 70 ± 38 pack-years smoking; 42% current smoking), 57 (26%) reported having any moderate to very severe AECOPD (m/s-AECOPD) while 66 (30%) had EHR documentation of m/s-AECOPD. However, 42% of those with EHR-identified m/s-AECOPD had none by self-report, and 33% of those who reported m/s-AECOPD had none by EHR, suggesting only moderate agreement (Cohens Kappa = 0.47 ± 0.07; P < 0.001). Nevertheless, self-reported and EHR-identified m/s-AECOPD events were both associated with higher BAL neutrophils (ß ± SEM: 3.0 ± 1.1 and 1.3 ± 0.5 per 10% neutrophil increase; P ≤ 0.018) and lymphocytes (0.9 ± 0.4 and 0.7 ± 0.3 per 10% lymphocyte increase; P ≤ 0.041). Exacerbation by either measure combined was associated with a larger estimated effect (3.7 ± 1.2 and 1.0 ± 0.5 per 10% increase in neutrophils and lymphocytes, respectively) but was not statistically significantly different compared to the self-report only approach. Among the 184 TEPS participants, there were fewer moderate to very severe respiratory exacerbations by self-report (n = 15 or 8%) or EHR-documentation (n = 9 or 5%), but a similar level of agreement as those with COPD was observed (Cohens Kappa = 0.38 ± 0.07; P < 0.001). DISCUSSION: While there is modest agreement between self-reported and EHR-identified m/s-AECOPD, events are missed by relying on either method alone. However, m/s-AECOPD quantified by self-report or health records is associated with BAL neutrophilia and lymphocytosis

In search of the ideal periosteal flap for bone non-union: the chimeric fibula-periosteal flap

Vascularized periosteal flaps may increase union rates in recalcitrant long bone non-union. The fibula-periosteal chimeric flap utilizes periosteum raised on an independent periosteal vessel. This allows the periosteum to be inset freely around the osteotomy site, thereby facilitating bone consolidation. Patients and Methods: 10 patients underwent fibula-periosteal chimeric flaps (2016 – 2022) at Canniesburn Plastic Surgery Unit, UK. Preceding non-union 18.6 months, with bone gap 7.5 cm. Patients underwent pre-operative CT angiography to identify periosteal branches. A case-control approach was used. Patients acted as their own controls with one osteotomy covered by the chimeric periosteal flap and one without, although in two patients both osteotomies were covered by a long periosteal flap. Results: A chimeric periosteal flap was used in 12 of 20 osteotomy sites. Periosteal flap osteotomies had a primary union rate of 100% (11/11) versus those without 28.6% (2/7) (p=0.0025). Union occurred in the chimeric periosteal flaps at 8.5 months versus 16.75 in the control group (p=0.023). 1 case excluded from primary analysis due to recurrent mycetoma. Number needed to treat = 2, indicating that 2 patients would require a chimeric periosteal flap to avoid one non-union. Survival curves with hazards ratio 4.1, equating to 4 times higher chance of union with periosteal flaps (log rank p=0.0016). Conclusions: The chimeric fibula-periosteal flap may increase consolidation rates in difficult cases of recalcitrant non-union. This elegant modification of the fibula flap uses periosteum that is normally discarded, and adds to accumulating data supporting the use of vascularised periosteal flaps in non-union

Differences in clinical presentation of primary open-angle glaucoma between African and European populations

PURPOSE: Primary open‐angle glaucoma (POAG) has been reported to occur more frequently in Africans, and to follow a more severe course compared to Europeans. We aimed to describe characteristics of POAG presentation and treatment across three ethnic groups from Africa and one from Europe. METHODS: We ascertained 151 POAG patients from South African Coloured (SAC) and 94 South African Black (SAB) ethnicity from a university hospital in South Africa. In Tanzania, 310 patients were recruited from a university hospital and a referral hospital. In the Netherlands, 241 patients of European ancestry were included. All patients were over 35 years old and had undergone an extensive ophthalmic examination. Patients were diagnosed according to the ISGEO criteria. A biogeographic ancestry analysis was performed to estimate the proportion of genetic African ancestry (GAA). RESULTS: The biogeographic ancestry analysis showed that the median proportion of GAA was 97.6% in Tanzanian, 100% in SAB, 34.2% in SAC and 1.5% in Dutch participants. Clinical characteristics at presentation for Tanzanians, SAB, SAC and Dutch participants, respectively: mean age: 63, 57, 66, 70 years (p < 0.001); visual acuity in the worse eye: 1.78, 1.78, 0.3, 0.3 LogMAR (p < 0.001); maximum intraocular pressure of both eyes: 36, 34, 29, 29 mmHg (p (anova)  < 0.001); maximum vertical cup to disc ratio (VCDR) of both eyes: 0.90, 0.90, 0.84, 0.83 (p < 0.001); mean central corneal thickness: 506, 487, 511, 528 μm (p < 0.001). Fourteen percent of Tanzanian patients presented with blindness (<3/60 Snellen) in the better eye in contrast to only 1% in the Dutch. CONCLUSION: In this multi‐ethnic comparative study, Sub‐Saharan Africans present at a younger age with lower visual acuity, higher IOP, larger VCDR, than SAC and Dutch participants. This indicates the more progressive and destructive course in Sub‐Saharan Africans

NRC Russian-English Machine Translation System for WMT 2016

We describe the statistical machine translation system developed at the National Research Council of Canada (NRC) for the Russian-English news translation task of the First Conference on Machine Translation (WMT 2016). Our submission is a phrase-based SMT system that tackles the morphological complexity of Russian through comprehensive use of lemmatization. The core of our lemmatization strategy is to use different views of Russian for different SMT components: word alignment and bilingual neural network language models use lemmas, while sparse features and reordering models use fully inflected forms. Some components, such as the phrase table, use both views of the source. Russian words that remain out-ofvocabulary (OOV) after lemmatization are transliterated into English using a statistical model trained on examples mined from the parallel training corpus. The NRC Russian-English MT system achieved the highest uncased BLEU and the lowest TER scores among the eight participants in WMT 2016

Does Industry-Driven Alcohol Marketing Influence Adolescent Drinking Behaviour? A Systematic Review.

AIM: To systematically review evidence on the influence of specific marketing components (Price, Promotion, Product attributes and Place of sale/availability) on key drinking outcomes (initiation, continuation, frequency and intensity) in young people aged 9–17. METHODS: MEDLINE, EMBASE, SCOPUS, PsychINFO, CINAHL and ProQuest were searched from inception to July 2015, supplemented with searches of Google Scholar, hand searches of key journals and backward and forward citation searches of reference lists of identified papers. RESULTS: Forty-eight papers covering 35 unique studies met inclusion criteria. Authors tended to report that greater exposure to alcohol marketing impacted on drinking initiation, continuation, frequency and intensity during adolescence. Nevertheless, 23 (66%) studies reported null results or negative associations, often in combination with positive associations, resulting in mixed findings within and across studies. Heterogeneity in study design, content and outcomes prevented estimation of effect sizes or exploration of variation between countries or age subgroups. The strength of the evidence base differed according to type of marketing exposure and drinking outcome studied, with support for an association between alcohol promotion (mainly advertising) and drinking outcomes in adolescence, whilst only two studies examined the relationship between alcohol price and the drinking behaviour of those under the age of 18. CONCLUSION: Despite the volume of work, evidence is inconclusive in all four areas of marketing but strongest for promotional activity. Future research with standardized measures is needed to build on this work and better inform interventions and policy responses

Clinical Grade Production of Wilms' Tumor-1 Loaded Cord Blood-Derived Dendritic Cells to Prevent Relapse in Pediatric AML After Cord Blood Transplantation

Hematopoietic cell transplantation (HCT) is a last resort, potentially curative treatment option for pediatric patients with refractory acute myeloid leukemia (AML). Cord blood transplantation (CBT) results in less relapses and less graft-versus-host disease when compared to other sources. Nevertheless, still more than half of the children die from relapses. We therefore designed a strategy to prevent relapses by inducing anti-AML immunity after CBT, using a CB-derived dendritic cell (CBDC) vaccine generated from CD34+ CB cells from the same graft. We here describe the optimization and validation of good manufacturing practice (GMP)-grade production of the CBDC vaccine. We show the feasibility of expanding low amounts of CD34+ cells in a closed bag system to sufficient DCs per patient for at least three rounds of vaccinations. The CBDCs showed upregulated costimulatory molecules after maturation and showed enhanced CCR7-dependent migration toward CCL19 in a trans-well migrations assay. CBDCs expressed Wilms' tumor 1 (WT1) protein after electroporation with WT1-mRNA, but were not as potent as CBDCs loaded with synthetic long peptides (peptivator). The WT1-peptivator loaded CBDCs were able to stimulate T-cells both in a mixed lymphocyte reaction as well as in an antigen-specific (autologous) setting. The autologous stimulated T-cells lysed not only the WT1+ cell line, but most importantly, also primary pediatric AML cells. Altogether, we provide a GMP-protocol of a highly mature CBDC vaccine, loaded with WT1 peptivator and able to stimulate autologous T-cells in an antigen-specific manner. Finally, these T-cells lysed primary pediatric AML demonstrating the competence of the CBDC vaccine strategy

Use of plasma DNA to predict mortality and need for intensive care in patients with abdominal pain

Background We investigated the value of plasma deoxyribonucleic acid concentrations in patients presenting with acute abdominal pain to predict need for intensive care or mortality. Methods Plasma deoxyribonucleic acid taken from patients with acute abdominal pain was analyzed for the β-globin gene using the quantitative polymerase chain reaction. The primary outcome measure was the combined 28-day mortality or admission to the intensive care unit. Results Of 287 consecutive patients with acute abdominal pain recruited, 12 patients were admitted to the intensive care unit and/or died. Median plasma DNA concentrations were higher in patients with cancer and major organ inflammation. Mean plasma DNA concentrations were three-fold higher in patients with systemic inflammatory response syndrome, five-fold higher in patients who died within 28 days, and eight-fold higher in patients admitted to the intensive care unit. The area under the receiver operator curve for plasma DNA concentrations and intensive care unit admission/mortality was 0.804. At a cut-off of 1100 GE/ml, the sensitivity was 67% (95%CI 35–90) and specificity was 89% (95%CI 84–92). At a cut-off of 175 GE/ml, the sensitivity was 100% (95%CI 73–100) and specificity was 30% (95%CI 25–36). Plasma DNA concentration predicted need for intensive care unit admission or death (adjusted odds ratio 1.4; P < 0.0001). Conclusions Plasma DNA may have a role in patients with acute abdominal pain as a marker for inflammation and cancer, and a predictor of intensive care unit admission/mortality