Intelligent image-based in situ single-cell isolation

Quantifying heterogeneities within cell populations is important for many fields including cancer research and neurobiology; however, techniques to isolate individual cells are limited. Here, we describe a high-throughput, non-disruptive, and cost-effective isolation method that is capable of capturing individually targeted cells using widely available techniques. Using high-resolution microscopy, laser microcapture microscopy, image analysis, and machine learning, our technology enables scalable molecular genetic analysis of single cells, targetable by morphology or location within the sample.Peer reviewe

CD62L (L-selectin) shedding for assessment of perioperative immune sensitivity in patients undergoing cardiac surgery with cardiopulmonary bypass

OBJECTIVE: To investigate the suitability of blood granulocyte and monocyte sensitivity, as measured by the quantity of different agonists required to induce CD62L shedding, for assessment of perioperative immune changes in patients undergoing cardiac surgery with cardiopulmonary bypass. METHODS: Patients scheduled for aortocoronary bypass grafting or for valve surgery were included in this prospective observational study. Blood samples were drawn before anesthesia induction, directly after surgery and 48 hours after anesthesia induction. We determined the concentration of two different inflammatory stimuli--lipoteichoic acid (LTA) and tumor necrosis factor alpha (TNF)--required to induce shedding of 50% of surface CD62L from blood granulocytes and monocytes. In parallel monocyte surface human leukocyte antigen (HLA)-DR, and plasma interleukin (IL)-8, soluble (s)CD62L, soluble (s)Toll-like receptor (TLR)-2 and ADAM17 quantification were used to illustrate perioperative immunomodulation. RESULTS: 25 patients were enrolled. Blood granulocytes and monocytes showed decreased sensitivity to the TLR 2/6 agonist Staphylococcus aureus LTA immediately after surgery (p = 0.001 and p = 0.004 respectively). In contrast, granulocytes (p = 0.01), but not monocytes (p = 0.057) displayed a decreased postoperative sensitivity to TNF. We confirmed the presence of a systemic inflammatory response and a decreased immune sensitivity in the post-surgical period by measuring significant increases in the perioperative plasma concentration of IL-8 (p </= 0.001) and sTLR (p = 0.004), and decreases in monocyte HLA-DR (p<0.001), plasma sCD62L (p </= 0.001). In contrast, ADAM17 plasma levels did not show significant differences over the observation period (p = 0.401). CONCLUSIONS: Monitoring granulocyte and monocyte sensitivity using the "CD62L shedding assay" in the perioperative period in cardiac surgical patients treated with the use of cardiopulmonary bypass reveals common changes in sensitivity to TLR2/6 ligands and to TNF stimulus. Further long-term follow-up studies will address the predictive value of these observations for clinical purposes

A comparison of nefazodone, the cognitive behavioral-analysis system of psychotherapy, and their combination for the treatment of chronic depression

Background Patients with chronic forms of major depression are difficult to treat, and the relative efficacy of medications and psychotherapy is uncertain. Methods We randomly assigned 681 adults with a chronic nonpsychotic major depressive disorder to 12 weeks of outpatient treatment with nefazodone (maximal dose, 600 mg per day), the cognitive behavioral-analysis system of psychotherapy (16 to 20 sessions), or both. At base line, all patients had scores of at least 20 on the 24-item Hamilton Rating Scale for Depression (indicating clinically significant depression). Remission was defined as a score of 8 or less at weeks 10 and 12. For patients who did not have remission, a satisfactory response was defined as a reduction in the score by at least 50 percent from base line and a score of 15 or less. Raters were unaware of the patients’ treatment assignments. Results Of the 681 patients, 662 attended at least one treatment session and were included in the analysis of response. The overall rate of response (both remission and satisfactory response) was 48 percent in both the nefazodone group and the psychotherapy group, as compared with 73 percent in the combined-treatment group (P Conclusions Although about half of patients with chronic forms of major depression have a response to short-term treatment with either nefazodone or a cognitive behavioral-analysis system of psychotherapy, the combination of the two is significantly more efficacious than either treatment alone

Replication Data for: Uncovering high-level corruption: Cross-national objective corruption risk indicators using public procurement data

Measuring high-level corruption is subject to extensive scholarly and policy interest with moderate progress in the last decade. We develop two objective proxy measures of high-level corruption in public procurement: single bidding in competitive markets and a composite score of tendering 'red flags'. Using official government data of 2.8 million contracts in 28 European countries in 2009-2014, we directly operationalize a common definition of corruption: unjustified restriction of access to public contracts to favour a selected bidder. Corruption indicators are calculated at the contract level, but produce aggregate indices consistent with well-established country-level indicators, and also validated by micro-level tests. Further data is published at http://digiwhist.eu/resources/data/

A HELYI GAZDASÁGFEJLESZTÉS GYAKORLATA A GYÖNGYÖSI KISTÉRSÉGBEN

A magyar területfejlesztési gyakorlatban, az utóbbi évtizedekben az a megközelítés volt az uralkodó, hogy az egyes térségek hiányzó erőforrásait kívülről érkező befektetők letelepítésével kívánták megoldani. Ennek köszönhetően a vidéki térségek kisebb jelentőségű erőforrásait gyakran részlegesen, vagy egyáltalán nem is hasznosították. A 2000-es évektől Magyarországon is külső és belső erőforrások kiegyensúlyozott felhasználását helyezték előtérbe, miközben fokozott elvárásként fogalmazódott meg a fenntarthatóság és a helyi közösségek szerepe a döntéshozatalban. A helyi gazdaságfejlesztés a legtöbb esetben tradicionális megoldásokban gondolkodik, amelyeket azonban modern társadalmi-gazdasági környezetben kell megvalósítani. A helyi gazdaságfejlesztés számos eszközére találunk példát az ország területén, azonban ezek gyakran csak pontszerűen fordulnak elő, míg egyes megoldások kifejezetten divatossá váltak. Vizsgálatunkban arra kerestük a választ, hogy a Gyöngyösi kistérség gazdaságfejlesztési gyakorlatában mennyire érvényesül ez a megközelítésmód, a helyi gazdaságfejlesztés mely eszközeinek alkalmazására találunk konkrét példákat. A mélyinterjú módszerével készült kutatás eredményei alapján kijelenthetjük, hogy a helyi gazdaságfejlesztés szemléletmódja egyre nagyobb teret nyer a Gyöngyösi kistérség területén is, azonban számos olyan eszköz van, ahol még csak tervekről, vagy induló projektekről beszélhetünk, több éves gyakorlati tapasztalatokról egyelőre nem. --------------------------------------------- In recent decades there was the dominant approach in the Hungarian regional development practice, that the missing resources of some regions was sought to solve by installing from outside investors. As a result of it, the marginal resources of the rural areas often were be recovered partially or not at all. From 2000’s in Hungary the balanced use of internal and external resources was come to the front, while the role of sustainability and local communities in decision-making was formulated as an increased expectations. Local economic development is a traditional solution in most cases, however, it should be achieved in modern social-economic environment. Many examples of the local economic development asstes can be found in Hungary, but they often occur only pointwise, and some solutions have become especially popular. In our study we examined to what extent this approach prevails the economic development practice in the micro region of Gyöngyös, which means of the local economic development application can be find specific examples. We can declare on the basis of the results of our interview research, that the approach of the local economic development grain ground more and more in the micro region of Gyöngyös as well, however, there are several tools, which are only plans or starting projects, we can’t talk about many years of practical experience yet

Technical Approach Determines Inflammatory Response after Surgical and Transcatheter Aortic Valve Replacement

To investigate the periprocedural inflammatory response in patients with isolated aortic valve stenosis undergoing surgical aortic valve replacement (SAVR) or transcatheter aortic valve implantation (TAVI) with different technical approaches.Patients were prospectively allocated to one of the following treatments: SAVR using conventional extracorporeal circulation (CECC, n = 47) or minimized extracorporeal circulation (MECC, n = 15), or TAVI using either transapical (TA, n = 15) or transfemoral (TF, n = 24) access. Exclusion criteria included infection, pre-procedural immunosuppressive or antibiotic drug therapy and emergency indications. We investigated interleukin (IL)-6, IL-8, IL-10, human leukocyte antigen (HLA-DR), white blood cell count, high-sensitivity C-reactive protein (hs-CRP) and soluble L-selectin (sCD62L) levels before the procedure and at 4, 24, and 48 h after aortic valve replacement. Data are presented for group interaction (p-values for inter-group comparison) as determined by the Greenhouse-Geisser correction.SAVR on CECC was associated with the highest levels of IL-8 and hs-CRP (p<0.017, and 0.007, respectively). SAVR on MECC showed the highest descent in levels of HLA-DR and sCD62L (both p<0.001) in the perioperative period. TA-TAVI showed increased intraprocedural concentration and the highest peak of IL-6 (p = 0.017). Significantly smaller changes in the inflammatory markers were observed in TF-TAVI.Surgical and interventional approaches to aortic valve replacement result in inflammatory modulation which differs according to the invasiveness of the procedure. As expected, extracorporeal circulation is associated with the most marked pro-inflammatory activation, whereas TF-TAVI emerges as the approach with the most attenuated inflammatory response. Factors such as the pre-treatment patient condition and the extent of myocardial injury also significantly affect inflammatory biomarker patterns. Accordingly, TA-TAVI is to be classified not as an interventional but a true surgical procedure, with inflammatory biomarker profiles comparable to those found after SAVR. Our study could not establish an obvious link between the extent of the periprocedural inflammatory response and clinical outcome parameters

NGS of Virus-Derived Small RNAs as a Diagnostic Method Used to Determine Viromes of Hungarian Vineyards

As virus diseases cannot be controlled by traditional plant protection methods, the risk of their spread have to be minimized on vegetatively propagated plants, such as grapevine. Metagenomic approaches used for virus diagnostics offer a unique opportunity to reveal the presence of all viral pathogens in the investigated plant, which is why their application can reduce the risk of using infected material for a new plantation. Here we used a special branch, deep sequencing of virus-derived small RNAs, of this high-throughput method for virus diagnostics, and determined viromes of vineyards in Hungary. With NGS of virus-derived small RNAs we could detect not only the viruses tested routinely, but also new ones, which had never been described in Hungary before. Virus presence did not correlate with the age of the plantation, moreover phylogenetic analysis of the identified virus isolates suggests that infections are mostly caused by the use of infected propagating material. Our results, validated by other molecular methods, raised further questions to be answered before this method can be introduced as a routine, reliable test for grapevine virus diagnostics