Extracellular vesicles (EVs) have great potential as biomarkers since their composition and concentration in biofluids are disease state dependent and their cargo can contain disease-related ...information. Large tumor-derived EVs (tdEVs, >1 μm) in blood from cancer patients are associated with poor outcome, and changes in their number can be used to monitor therapy effectiveness. Whereas, small tumor-derived EVs (<1 μm) are likely to outnumber their larger counterparts, thereby offering better statistical significance, identification and quantification of small tdEVs are more challenging. In the blood of cancer patients, a subpopulation of EVs originate from tumor cells, but these EVs are outnumbered by non-EV particles and EVs from other origin. In the Dutch NWO Perspectief Cancer-ID program, we developed and evaluated detection and characterization techniques to distinguish EVs from non-EV particles and other EVs. Despite low signal amplitudes, we identified characteristics of these small tdEVs that may enable the enumeration of small tdEVs and extract relevant information. The insights obtained from Cancer-ID can help to explore the full potential of tdEVs in the clinic.
Abstract Study Objective To determine whether spinal anesthesia blunts surgical stress reactions and results in less perioperative hyperglycemia. Design Prospective, randomized controlled study. ...Setting Operating room of a university hospital. Patients 68 adult, nondiabetic (n=40) and diabetic (n=28), ASA physical status 1, 2, and 3 patients patients undergoing elective total hip replacement. Interventions General or spinal anesthesia was administered. Measurements Blood HbA1C was measured preoperatively to identify patients with undiagnosed diabetes. Glucose levels were checked preoperatively, then immediately after, and one hour after surgery. A conventional glucose control protocol was used, where insulin was given when blood glucose concentrations exceeded 250 mg/dL. Main Results Preoperative glucose levels in general and spinal anesthesia patients were comparable and not significantly different in nondiabetic and diabetic patients. At the end of surgery and one hour after surgery, glucose levels were significantly higher in patients undergoing general anesthesia compared with baseline values in both diabetic and nondiabetic patients ( P < 0.05). In nondiabetic and diabetic patients, a significant increase in glucose level was found in patients undergoing general anesthesia versus spinal anesthesia ( P < 0.05). In patients receiving spinal anesthesia, glucose levels remained stable. Two diabetic patients undergoing general anesthesia received insulin. Conclusion Spinal anesthesia attenuates the hyperglycemic response to surgical stimuli in diabetics and nondiabetic patients.
In the randomized phase II DREAMM-2 study, single-agent belantamab mafodotin demonstrated deep and durable responses and a manageable safety profile in triple-class refractory relapsed/refractory ...multiple myeloma (RRMM). We present patient-reported outcomes (PROs) from this study for patients treated with the approved dose of belantamab mafodotin (2.5 mg/kg q3w). Disease and treatment-related symptoms, health-related quality of life (HRQOL), functioning, and patient-reported ocular changes were assessed using questionnaires (European Organisation for Research and Treatment of Cancer Quality of Life questionnaires EORTC-QLQ-C30 and EORTC-QLQ-MY20, Ocular Surface Disease Index OSDI, and the National Eye Institute Visual Functioning Questionnaire 25 NEI VFQ-25) at baseline, during treatment (every 3 or 6 weeks), and at the end of treatment (EOT). Eye examinations were conducted at baseline, prior to each treatment cycle, and at EOT. Patients reported ocular symptoms in the OSDI and NEI VFQ-25 questionnaires, with the median time to worst severity of 45 to 64 days depending on symptoms considered. Some limitations in driving and reading were reported. Ocular symptoms were improved and median time to recovery was 23.5 to 44.0 days. EORTC-QLQ-C30 data suggest core MM symptoms (including fatigue and pain), overall HRQOL, and patient functioning were maintained while patients continued belantamab mafodotin treatment, even if meaningful worsening of vision-related symptoms occurred. These PRO results, together with the clinical efficacy of belantamab mafodotin, support its use in patients with RRMM and further evaluation of its use at earlier lines of therapy.
Objective In squamous cell carcinoma of the pharynx and larynx, NOTCH1 downstream signaling has been shown to be activated. The NOTCH1 signaling pathway has not been examined in detail for sinonasal ...squamous cell carcinomas (SNSCCs). The aim of this study was to evaluate NOTCH1 signaling by mRNA expression analysis and to examine the occurrence of NOTCH1 mutations in SNSCC. Study Design In a retrospective study, we analyzed tissues from 44 SNSCCs and 56 head and neck squamous cell carcinomas (HNSCCs) at other locations. Expression of NOTCH1, NOTCH3, HES1, HEY1, and JAG1 mRNA were measured by using quantitative real-time polymerase chain reaction (q-rtPCR). In SNSCC, NOTCH1 mutations were evaluated with sequencing of seven selected exons. Results Expression of NOTCH1, HEY1, and JAG1 at the mRNA level were significantly higher in tumor tissue compared with normal tissue. In SNSCC, the subgroup of patients with high expression (5th quintile) of HES1 mRNA was associated with better survival ( P = .04); however these patients with high expression of HES1 mRNA had also a more favorable tumor stage and grade and more unfavorable resections representing potential confounders. Conclusions Key components of NOTCH1 are upregulated at the mRNA level in HNSCCs. The mechanism, clinical significance, and potential therapeutic options should therefore be further evaluated.
The blood-brain barrier (BBB) prevents entry of neurotoxic substances but also that of drugs into the brain. Here, the paracellular barrier is formed by tight junctions (TJs) with claudin-5 (Cldn5) ...being the main sealing constituent. Transient BBB opening by targeting Cldn5 could improve paracellular drug delivery. The non-toxic C-terminal domain of Clostridium perfringens enterotoxin (cCPE) binds to a subset of claudins, e.g., Cldn3, -4. Structure-based mutagenesis was used to generate Cldn5-binding variants (cCPE-Y306W/S313H and cCPE-N218Q/Y306W/S313H). These cCPE-variants were tested for transient TJ opening using multiple in vitro BBB models: Primary porcine brain endothelial cells, coculture of primary rat brain endothelial cells with astrocytes and mouse cerebEND cells. cCPE-Y306W/S313H and cCPE-N218Q/Y306W/S313H but neither cCPE-wt nor cCPE-Y306A/L315A (not binding to claudins) decreased transendothelial electrical resistance in a concentration-dependent and reversible manner. Furthermore, permeability of carboxyfluorescein (with size of CNS drugs) was increased. cCPE-Y306W/S313H but neither cCPE-wt nor cCPE-Y306A/L315A bound to Cldn5-expressing brain endothelial cells. However, freeze-fracture EM showed that cCPE-Y306W/S313H did not cause drastic TJ breakdown. In sum, Cldn5-binding cCPE-variants enabled mild and transient opening of brain endothelial TJs. Using reliable in vitro BBB models, the results demonstrate that cCPE-based biologics designed to bind Cldn5 improve paracellular drug delivery across the BBB.
•Clostridium perfringens enterotoxin-based claudin-5 modulators were generated.•They improve blood brain barrier permeability in size-selective and reversible manner.•They provide novel biologicals for drug delivery across blood brain barrier.
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The photoproduction of
and
mesons off carbon and niobium nuclei has been measured as a function of the meson momentum for incident photon energies of 1.2-2.9GeV at the electron accelerator ELSA. ...The mesons have been identified via the
and
decays, respectively, registered with the CBELSA/TAPS detector system. From the measured meson momentum distributions the momentum dependence of the transparency ratio has been determined for both mesons. Within a Glauber analysis the in-medium
and
widths and the corresponding absorption cross sections have been deduced as a function of the meson momentum. The results are compared to recent theoretical predictions for the in-medium
width and
-N absorption cross sections. The energy dependence of the imaginary part of the
- and
-nucleus optical potential has been extracted. The finer binning of the present data compared to the existing data allows a more reliable extrapolation towards the production threshold. The modulus of the imaginary part of the
-nucleus potential is found to be about three times smaller than recently determined values of the real part of the
-nucleus potential, which makes the
meson a suitable candidate for the search for meson-nucleus bound states. For the
meson, the modulus of the imaginary part near threshold is comparable to the modulus of the real part of the potential. As a consequence, only broad structures can be expected, which makes the observation of
mesic states very difficult experimentally.
Tight junctions regulate paracellular permeability size and charge selectively. Models have been proposed for the molecular architecture of tight junction strands and paracellular channels. However, ...they are not fully consistent with experimental and structural data. Here, we analysed the architecture of claudin-based tight junction strands and channels by cellular reconstitution of strands, structure-guided mutagenesis, in silico protein docking and oligomer modelling. Prototypic channel- (Cldn10b) and barrier-forming (Cldn3) claudins were analysed. Förster resonance energy transfer (FRET) assays indicated multistep claudin polymerisation, starting with cis-oligomerization specific to the claudin subtype, followed by trans-interaction-triggered cis-polymerisation. Alternative protomer interfaces were modelled in silico and tested by cysteine-mediated crosslinking, confocal- and freeze fracture EM-based analysis of strand formation. The analysed claudin mutants included also mutations causing the HELIX syndrome. The results indicated that protomers in Cldn10b and Cldn3 strands form similar antiparallel double rows, as has been suggested for Cldn15. Mutually stabilising ‐hydrophilic and hydrophobic ‐ cis- and trans-interfaces were identified that contained novel key residues of extracellular segments ECS1 and ECS2.
Hydrophobic clustering of the flexible ECS1 β1β2 loops together with ECS2–ECS2 trans-interaction is suggested to be the driving force for conjunction of tetrameric building blocks into claudin polymers. Cldn10b and Cldn3 are indicated to share this polymerisation mechanism. However, in the paracellular centre of tetramers, electrostatic repulsion may lead to formation of pores (Cldn10b) and electrostatic attraction to barriers (Cldn3). Combining in vitro data and in silico modelling, this study improves mechanistic understanding of paracellular permeability regulation by elucidating claudin assembly and its pathologic alteration as in HELIX syndrome.
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•Multistep polymerisation of claudins is driven by mutual stabilisation of cis- and trans-interfaces.•Claudin-3 and -10b differ in oligomerization but share similar face-to-face double-row architecture in strands.•Strand polymerisation occurs independent of channel versus barrier conformation.•Structural keys for claudin assembly determining paracellular permeability were identified.
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