Circulating microRNAs (miRNAs) have become increasingly popular biomarker candidates in various diseases. However, heparin-based anticoagulants might affect the detection of target miRNAs in blood ...samples during quantitative polymerase chain reaction (qPCR)-based analysis of miRNAs involving RNA extraction, cDNA synthesis and the polymerase catalyzed reaction. Because low-molecular-weight heparins (LMWH) are widely used in routine healthcare, we aimed to investigate whether a prophylactic dose of the LMWH tinzaparin influences qPCR-based quantification of circulating miRNAs. A total of 30 subjects were included: 16 fracture patients with tinzaparin treatment and 14 non-fracture controls without anticoagulation therapy. To control for the effect of tinzaparin on miRNA analysis an identical concentration of synthetic miRNAs was added to plasma, isolated RNA and prepared complementary DNA (cDNA) from all samples in both groups. No significant difference was observed for cDNA synthesis or qPCR when comparing tinzaparin-treated patients with untreated controls. Among the tinzaparin-treated patients, plasma levels of six endogenous miRNAs (hsa-let-7i-5p, hsa-miR-30e-5p, hsa-miR-222-3p, hsa-miR-1-3p, hsa-miR-133a-3p, hsa-miR-133b) were measured before and one to six hours after a subcutaneous injection of tinzaparin 4500IU. No significant effect was observed for any of the investigated miRNAs. A prophylactic dose of 4500IU tinzaparin does not seem to affect cDNA synthesis or qRT-PCR-based quantification of circulating miRNAs.
Circulating microRNAs (miRNAs) have become increasingly popular biomarker candidates in various diseases. However, heparin-based anticoagulants might affect the detection of target miRNAs in blood ...samples during quantitative polymerase chain reaction (qPCR)-based analysis of miRNAs involving RNA extraction, cDNA synthesis and the polymerase catalyzed reaction. Because low-molecular-weight heparins (LMWH) are widely used in routine healthcare, we aimed to investigate whether a prophylactic dose of the LMWH tinzaparin influences qPCR-based quantification of circulating miRNAs. A total of 30 subjects were included: 16 fracture patients with tinzaparin treatment and 14 non-fracture controls without anticoagulation therapy. To control for the effect of tinzaparin on miRNA analysis an identical concentration of synthetic miRNAs was added to plasma, isolated RNA and prepared complementary DNA (cDNA) from all samples in both groups. No significant difference was observed for cDNA synthesis or qPCR when comparing tinzaparin-treated patients with untreated controls. Among the tinzaparin-treated patients, plasma levels of six endogenous miRNAs (hsa-let-7i-5p, hsa-miR-30e-5p, hsa-miR-222-3p, hsa-miR-1-3p, hsa-miR-133a-3p, hsa-miR-133b) were measured before and one to six hours after a subcutaneous injection of tinzaparin 4500IU. No significant effect was observed for any of the investigated miRNAs. A prophylactic dose of 4500IU tinzaparin does not seem to affect cDNA synthesis or qRT-PCR-based quantification of circulating miRNAs.
This study was regarding the waterjets in the water atomization process. This is because the understanding of the waterjets is not complete and with a greater understanding the production of metal ...powder could be improved. The waterjets were going to be categorized according to their wave function, size and distribution of the droplets and the three regimes that Höganäs had divided up the jets into was also analyzed. The three regimes depend on the jets characteristics and the regimes are the transparent, milky and the droplet jet. The purpose was to get a better understanding of the correlation between velocity, temperature, waves, size and distribution of the droplets in a 50 cm long waterjet. The method to enhance the understanding of this project was to first do theoretical solution with the help of fluid dynamics. Weber, Reynolds and Ohnesorge number were calculated and evaluated to get a better understanding of the waterjet. Secondly, experiments were conducted where a waterjet with different nozzles and temperatures was filmed with a highspeed camera and the videos were analyzed with the help of a software package called ImageJ. The results show the correlation between increasing temperature and decreasing droplet size and a less cohesive waterjet core. The conclusion from the study was that with the help of temperature one can help control the droplet size.
Denna studie angår vattenstrålar i en vattenatomiserings process vid tillverkning av metallpulver och en bättre förståelse skulle förbättra tillverkningen av metallpulver. Vattenstrålen skulle kartläggas enligt dess vågfunktion, storlek och spridning av dropparna och de tre regionerna som Höganäs har delat upp vattenstrålen i skulle analyseras. Regionerna är beroende av strålens karaktäristiska utseenden vid olika delare av vattenstrålen och är genomskinlig, mjölkig och dropp stråle. Syftet med studien var att få en bättre förståelse av sambandet mellan hastighet, temperatur, vågor, storlek och spridning av dropparna i en 50 cm lång vattenstråle. Metoden som användes för att öka förståelsen av vattenstrålen var först en teoretisk del med hjälp av fluidmekanik. Weber, Reynolds och Ohnesorge tal beräknades och utvärderades för att ge en teoretisk förståelse för vattenstrålen. Sedan utfördes vattenflödesexperiment, där vattenstrålen filmades med olika munstycken och temperaturer med en höghastighetskamera och videon analyserades med hjälp av mjukvaran kallad ImageJ. Resultaten visar ett samband mellan ökad temperatur och minskad droppstorlek och en mer uppbruten kärna av vattenstrålen. Slutsatsen från studien var att man med hjälp av temperaturen kan reglera droppstorleken.
Efter revisionspliktens slopande år 2010 har fler förenklingar för småföretag blivit föreslagna angående revision och redovisning. En av dessa är slopandet av de jävsbestämmelser som är relaterade ...till revision för småföretag, nämligen redovisnings-, medelsförvaltnings- och byråjävet. Förslaget avsåg att förenkla för småföretag samt att öka möjligheterna för revisionsbyråers så kallade kombiuppdrag. Denna debatt har lett till ett lagförslag om ett slopande av dessa jävsregler vilket sedermera blivit avslaget med motiveringen att det skulle äventyra revisionens oberoende. Vi vill med denna studie undersöka vilka positiva och negativa aspekter som ett slopande av redovisningsjävsreglerna skulle resultera i samt vilka som skulle gagnas mest av en sådan förändring. De teorier som används som grund för denna kvalitativa studie är Principal-agent teorin, Oberoende, Legitimitetsteorin samt Intressentmodellen, inom Principal-agent teorin finns en delteori som heter Informationsasymmetrier. Dessa teorier ger oss som forskare en god kunskapsgrund samtidigt som dessa används som underlag för intervjumanualerna och för den senare analysen av empirin. I syfte att studera vårt ämne samt försöka besvara våra problemformuleringar och syfte har vi genomfört djupintervjuer med ett målstyrt urval. Dessa intervjuer genomfördes utifrån ett semistrukturerat tillvägagångsätt i syfte att uppnå en mer konversationsliknande intervju samt att låta respondenterna utveckla sina egna åsikter. Dessa respondenter utgjordes av en egen företagare, en revisor, en redovisningskonsult, en banktjänsteman samt två forskare. Vi valde att intervjua personer från olika yrken i syfte att få olika infallsvinklar på denna fråga och på det sättet få en bättre uppfattning och kunskap om ämnet. Utifrån den empiri som vi insamlat kan vi med hjälp av vår teoretiska referensram analysera denna subjektivt. Utifrån de utförda intervjuerna samt teori kan vi dra slutsatsen att denna regeländring skulle innebära positiva, negativa samt oförändrade aspekter. De övervägande positiva aspekterna utgörs av ökad effektivitet internt för revisionsbyråer, praktisk förenkling för småföretag, ökade affärsmöjligheter för revisionsbyråer samt effektivare rådgivning för småföretag. De negativa aspekterna utgörs av att revisionens kvalitetsstämpel för småföretag försvinner, minskade affärsmöjligheter för redovisningskonsultbyråer, sämre skatteunderlag för skattemyndigheten och att revisionsbyråerna tappar en del av sin professionalitet. Eftersom redovisningsjävet enbart berör små företag vars nytta med revision främst är intern istället för extern anses de negativa aspekterna vara av mindre vikt än de positiva. Således är slutsatsen att slopandet av redovisningsjävsreglerna överlag skulle vara positivt.
Background
Achieving a pathologic complete response (pCR) with neoadjuvant chemotherapy (NAC) in patients with muscle‐invasive bladder cancer (MIBC) has been associated with improved overall survival ...(OS). This study was aimed at evaluating the impact of pathologic downstaging (pDS; ie, a pT stage at least 1 stage lower than the pre‐NAC cT stage) on the OS of patients with MIBC treated with NAC.
Methods
The Retrospective International Study of Cancers of the Urothelial Tract (RISC) and the National Cancer Database (NCDB) were queried for cT2‐4N0M0 patients treated with NAC. A multivariable Cox model including either pDS or pCR was generated. A nested model was built to evaluate the added value of pDS (excluding patients achieving a pCR) to a model including pCR alone. C indices were computed to assess discrimination. NCDB was used for validation. The treatment effect of NAC versus cystectomy alone in achieving pDS was estimated through an inverse probability–weighted regression adjustment.
Results
Overall, 189 and 2010 patients from the RISC and NCDB cohorts, respectively, were included; pDS and pCR were achieved by 33% and 35% and by 20% and 15% in RISC and NCDB, respectively. In both data sets, pDS and pCR were associated with better OS and C indices. Adding pDS excluding pCR to the model with pCR fit the data better (likelihood ratio, P = .019 for RISC and P < .001 for NCDB), and it yielded better discrimination (incremental C index, 4.2 for RISC and 1.6 for NCDB). The treatment effect of NAC in achieving pDS was 2.07‐fold (P < .001) in comparison with cystectomy alone.
Conclusions
A decrease of at least 1 stage from the cT stage to the pT stage is associated with improved OS in patients with MIBC treated with NAC.
Using 2 different cohorts, this study demonstrates that a decrease in tumor size, even without complete disappearance, is associated with better survival than no response in patients with muscle‐invasive bladder cancer treated with neoadjuvant chemotherapy.
One hallmark of multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE) is infiltration of leukocytes into the CNS, where chemokines and their receptors play a major mediatory ...role. CX3CR1 is a chemokine receptor involved in leukocyte adhesion and migration and hence a mediator of immune defense reactions. The role of CX3CR1 in MS and EAE pathogenesis however remains to be fully assessed. Here, we demonstrate CX3CR1 mRNA expression on inflammatory cells within active plaque areas in MS brain autopsies. To test whether blocking CNS infiltration of peripheral leukocytes expressing CX3CR1 would be a suitable treatment strategy for MS, we developed a selective, high-affinity inhibitor of CX3CR1 (AZD8797). The compound is active outside the CNS and AZD8797 treatment in Dark Agouti rats with myelin oligodendrocyte glycoprotein-induced EAE resulted in reduced paralysis, CNS pathology, and incidence of relapses. The compound is effective when starting treatment before onset, as well as after the acute phase. This treatment strategy is mechanistically similar to, but more restricted than, current very late antigen-4–directed approaches that have significant side effects. We suggest that blocking CX3CR1 on leukocytes outside the CNS could be an alternative approach to treat MS.
The global surge in demand for metals such as cobalt and nickel has created unprecedented interest in deep-sea habitats with mineral resources. The largest area of activity is a 6 million km2 region ...known as the Clarion-Clipperton Zone (CCZ) in the central and eastern Pacific, regulated by the International Seabed Authority (ISA). Baseline biodiversity knowledge of the region is crucial to effective management of environmental impact from potential deep-sea mining activities, but until recently this has been almost completely lacking. The rapid growth in taxonomic outputs and data availability for the region over the last decade has allowed us to conduct the first comprehensive synthesis of CCZ benthic metazoan biodiversity for all faunal size classes. Here we present the CCZ Checklist, a biodiversity inventory of benthic metazoa vital to future assessments of environmental impacts. An estimated 92% of species identified from the CCZ are new to science (436 named species from a total of 5,578 recorded). This is likely to be an overestimate owing to synonyms in the data but is supported by analysis of recent taxonomic studies suggesting that 88% of species sampled in the region are undescribed. Species richness estimators place total CCZ metazoan benthic diversity at 6,233 (+/−82 SE) species for Chao1, and 7,620 (+/−132 SE) species for Chao2, most likely representing lower bounds of diversity in the region. Although uncertainty in estimates is high, regional syntheses become increasingly possible as comparable datasets accumulate. These will be vital to understanding ecological processes and risks of biodiversity loss.
•We provide the first checklist for the Clarion Clipperton Zone (CCZ) metazoan fauna•5,142 unnamed species (informal names) are recorded from the CCZ•Total estimates of species richness range from >6,000–>8,000•An estimated 88%–92% of species in the CCZ region in total are undescribed
Species-level biodiversity information is key to understanding ecosystems and tracking environmental impacts. Rabone et al. provide the first checklist (436 species) and total species estimates (>6,000–>8,000) for the world’s largest mineral exploration region, the CCZ. Estimates provide a baseline to build biodiversity knowledge at a regional scale.
Multiple myeloma (MM) is at present an incurable malignancy, characterized by apoptosis-resistant tumor cells. Interferon (IFN) treatment sensitizes MM cells to Fas-induced apoptosis and is ...associated with an increased activation of Signal transducer and activator of transcription (Stat)1. The role of Stat1 in MM has not been elucidated, but Stat1 has in several studies been ascribed a pro-apoptotic role. Conversely, IL-6 induction of Stat3 is known to confer resistance to apoptosis in MM.
To delineate the role of Stat1 in IFN mediated sensitization to apoptosis, sub-lines of the U-266-1970 MM cell line with a stable expression of the active mutant Stat1C were utilized. The influence of Stat1C constitutive transcriptional activation on endogenous Stat3 expression and activation, and the expression of apoptosis-related genes were analyzed. To determine whether Stat1 alone would be an important determinant in sensitizing MM cells to apoptosis, the U-266-1970-Stat1C cell line and control cells were exposed to high throughput compound screening (HTS).
To explore the role of Stat1 in IFN mediated apoptosis sensitization of MM, we established sublines of the MM cell line U-266-1970 constitutively expressing the active mutant Stat1C. We found that constitutive nuclear localization and transcriptional activity of Stat1 was associated with an attenuation of IL-6-induced Stat3 activation and up-regulation of mRNA for the pro-apoptotic Bcl-2 protein family genes Harakiri, the short form of Mcl-1 and Noxa. However, Stat1 activation alone was not sufficient to sensitize cells to Fas-induced apoptosis. In a screening of > 3000 compounds including bortezomib, dexamethasone, etoposide, suberoylanilide hydroxamic acid (SAHA), geldanamycin (17-AAG), doxorubicin and thalidomide, we found that the drug response and IC50 in cells constitutively expressing active Stat1 was mainly unaltered.
We conclude that Stat1 alters IL-6 induced Stat3 activity and the expression of pro-apoptotic genes. However, this shift alone is not sufficient to alter apoptosis sensitivity in MM cells, suggesting that Stat1 independent pathways are operative in IFN mediated apoptosis sensitization.
Although guidelines exist for advanced and variant bladder cancer management, evidence is limited/conflicting in some areas and the optimal approach remains controversial.
To bring together a large ...multidisciplinary group of experts to develop consensus statements on controversial topics in bladder cancer management.
A steering committee compiled proposed statements regarding advanced and variant bladder cancer management which were assessed by 113 experts in a Delphi survey. Statements not reaching consensus were reviewed; those prioritised were revised by a panel of 45 experts prior to voting during a consensus conference.
Online Delphi survey and consensus conference.
The European Association of Urology (EAU), the European Society for Medical Oncology (ESMO), experts in bladder cancer management.
Statements were ranked by experts according to their level of agreement: 1-3 (disagree), 4-6 (equivocal), and 7-9 (agree). A priori (level 1) consensus was defined as ≥70% agreement and ≤15% disagreement, or vice versa. In the Delphi survey, a second analysis was restricted to stakeholder group(s) considered to have adequate expertise relating to each statement (to achieve level 2 consensus).
Overall, 116 statements were included in the Delphi survey. Of these statements, 33 (28%) achieved level 1 consensus and 49 (42%) achieved level 1 or 2 consensus. At the consensus conference, 22 of 27 (81%) statements achieved consensus. These consensus statements provide further guidance across a broad range of topics, including the management of variant histologies, the role/limitations of prognostic biomarkers in clinical decision making, bladder preservation strategies, modern radiotherapy techniques, the management of oligometastatic disease, and the evolving role of checkpoint inhibitor therapy in metastatic disease.
These consensus statements provide further guidance on controversial topics in advanced and variant bladder cancer management until a time when further evidence is available to guide our approach.
This report summarises findings from an international, multistakeholder project organised by the EAU and ESMO. In this project, a steering committee identified areas of bladder cancer management where there is currently no good-quality evidence to guide treatment decisions. From this, they developed a series of proposed statements, 71 of which achieved consensus by a large group of experts in the field of bladder cancer. It is anticipated that these statements will provide further guidance to health care professionals and could help improve patient outcomes until a time when good-quality evidence is available.
There is a growing interest in the exploitation of deep-sea mineral deposits, particularly on the abyssal seafloor of the central Pacific Clarion-Clipperton Zone (CCZ), which is rich in polymetallic ...nodules. In order to effectively manage potential exploitation activities, a thorough understanding of the biodiversity, community structure, species ranges, connectivity, and ecosystem functions across a range of scales is needed. The benthic megafauna plays an important role in the functioning of deep-sea ecosystems and represents an important component of the biodiversity. While megafaunal surveys using video and still images have provided insight into CCZ biodiversity, the collection of faunal samples is needed to confirm species identifications to accurately estimate species richness and species ranges, but faunal collections are very rarely carried out. Using a Remotely Operated Vehicle, 55 specimens of benthic megafauna were collected from seamounts and abyssal plains in three Areas of Particular Environmental Interest (APEI 1, APEI 4, and APEI 7) at 3100-5100 m depth in the western CCZ. Using both morphological and molecular evidence, 48 different morphotypes belonging to five phyla were found, only nine referrable to known species, and 39 species potentially new to science. This work highlights the need for detailed taxonomic studies incorporating genetic data, not only within the CCZ, but in other bathyal, abyssal, and hadal regions, as representative genetic reference libraries that could facilitate the generation of species inventories.
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