Kinetics of endoglucanase and endoxylanase uptake by soybean seeds

The uptakes of endoglucanase and endoxylanase by soybean were studied by measuring the fluorescent intensity of a carboxyfluorescein. The disappearance rates of these enzymes from solution were compared with predictions from different models. The Freundlich model provided the best fit for our data. Enzyme concentration in the solution did not change significantly after 11 d, and therefore it was assumed that equilibrium was reached. Average mass transfer coefficient was calculated for both endoglucanase and endoxylanase from the plot of the rate of uptake against (C(l av) -Cl*) and a value of 4 x 10(-5) ms(-1) was obtained. K value was used to calculate the effective diffusivities of the two enzymes assuming a slice. It is found that K was asymptotic with long residence times, and a value of 4.8 x 10(-10) m2 s(-1) was obtained.     

HCO3 increment in arterial line can reveal significant vascular access recirculation in high-flux hemodialysis: a preliminary report

We report a new and simple way that can reveal the presence of vascular access recirculation (VAR) in patients undergoing hemodialysis (HD). Acid-base and blood gas parameters (pH, pO(2), pCO(2), and HCO(3)) were measured in blood samples drawn from an arterial fistula needle before the initiation of HD and from arterial and venous lines simultaneously 5 min later, in 31 patients (group A). Vascular access recirculation was measured using the glucose infusion test (GIT) immediately after the withdrawal of the 5-min samples. The same study was repeated in 30 patients in whom HD lines were reversed (group B). A comparison with baseline (predialysis) values of an analysis of the arterial line in group A at 5 min revealed that pCO(2) increased by 1.14+/-2.5 mmHg and HCO(3) by 0.6+/-0.6 mM/L (p<0.02 and p<0.00001, respectively). The corresponding pO(2) and pH values did not show significant differences. Glucose infusion test at 5 min (GITa) was -0.058+/-0.03%. A comparison with baseline (predialysis) values of an analysis of the arterial line in group B at 5 min revealed that pCO(2) increased by 7.7+/-3.5 mmHg and HCO(3) by 2.9+/-1.0 mM/L (p<0.000001 in each case). The pH level was significantly lower in comparison with baseline values (p<0.00001), while pO(2) did not show a significant difference. Glucose infusion test at 5 min (GITb) was 12.0+/-6.1% (p<0.000001 in comparison with GITa values). Clinically significant VAR was defined as HCO(3) increment >1.8 mM/L, based on the receiver-operating characteristics curve, which showed a threshold value of HCO(3) increment >1.8 mmol/L as a predictor of GIT recirculation. Five minutes after the initiation of high-flux HD with a 0 ultrafiltration rate, there is a small increment in arterial HCO(3) values relative to predialysis values. Clinically significant VAR is present when this increment is higher than 1.8 mM/L.     

Efficient removal of microcystin-LR by UV-C/H₂O₂ in synthetic and natural water samples

The destruction of the commonly found cyanobacterial toxin, microcystin-LR (MC-LR), in surface waters by UV-C/H₂O₂ advanced oxidation process (AOP) was studied. Experiments were carried out in a bench scale photochemical apparatus with low pressure mercury vapor germicidal lamps emitting at 253.7 nm. The degradation of MC-LR was a function of UV fluence. A 93.9% removal with an initial MC-LR concentration of 1 μM was achieved with a UV fluence of 80 mJ/cm² and an initial H₂O₂ concentration of 882 μM. When increasing the concentration of MC-LR only, the UV fluence-based pseudo-first order reaction rate constant generally decreased, which was probably due to the competition between by-products and MC-LR for hydroxyl radicals. An increase in H₂O₂ concentration led to higher removal efficiency; however, the effect of HO scavenging by H₂O₂ became significant for high H₂O₂ concentrations. The impact of water quality parameters, such as pH, alkalinity and the presence of natural organic matter (NOM), was also studied. Field water samples from Lake Erie, Michigan and St. Johns River, Florida were employed to evaluate the potential application of this process for the degradation of MC-LR. Results showed that the presence of both alkalinity (as 89.6-117.8 mg CaCO₃/L) and NOM (as ∼2 to ∼9.5 mg/L TOC) contributed to a significant decrease in the destruction rate of MC-LR. However, a final concentration of MC-LR bellow the guideline value of 1 μg/L was still achievable under current experimental conditions when an initial MC-LR concentration of 2.5 μg/L was spiked into those real water samples.     

Extracellular Vesicles in Myeloid Neoplasms

Myeloid neoplasms arise from malignant primitive cells, which exhibit growth advantage within the bone marrow microenvironment (BMM). The interaction between these malignant cells and BMM cells is critical for the progression of these diseases. Extracellular vesicles (EVs) are lipid bound vesicles secreted into the extracellular space and involved in intercellular communication. Recent studies have described RNA and protein alterations in EVs isolated from myeloid neoplasm patients compared to healthy controls. The altered expression of various micro-RNAs is the best-described feature of EVs of these patients. Some of these micro-RNAs induce growth-related pathways such as AKT/mTOR and promote the acquisition of stem cell-like features by malignant cells. Another well-described characteristic of EVs in myeloid neoplasms is their ability to suppress healthy hematopoiesis either via direct effect on healthy CD34+ cells or via alteration of the differentiation of BMM cells. These results support a role of EVs in the pathogenesis of myeloid neoplasms. mainly through mediating the interaction between malignant and BMM cells, and warrant further study to better understand their biology. In this review, we describe the reported alterations of EV composition in myeloid neoplasms and the recent discoveries supporting their involvement in the development and progression of these diseases.     

A Degron Blocking Strategy Towards Improved CRL4CRBN Recruiting PROTAC Selectivity**

Small molecules inducing protein degradation are important pharmacological tools to interrogate complex biology and are rapidly translating into clinical agents. However, to fully realise the potential of these molecules, selectivity remains a limiting challenge. Herein, we addressed the issue of selectivity in the design of CRL4^CRBN recruiting PROteolysis TArgeting Chimeras (PROTACs). Thalidomide derivatives used to generate CRL4^CRBN recruiting PROTACs have well described intrinsic monovalent degradation profiles by inducing the recruitment of neo-substrates, such as GSPT1, Ikaros and Aiolos. We leveraged structural insights from known CRL4^CRBN neo-substrates to attenuate and indeed remove this monovalent degradation function in well-known CRL4^CRBN molecular glues degraders, namely CC-885 and Pomalidomide. We then applied these design principles on a previously published BRD9 PROTAC (dBRD9-A) and generated an analogue with improved selectivity profile. Finally, we implemented a computational modelling pipeline to show that our degron blocking design does not impact PROTAC-induced ternary complex formation. We believe that the tools and principles presented in this work will be valuable to support the development of targeted protein degradation.