Arylazopyrazole AAP1742 Inhibits CDKs and Induces Apoptosis in Multiple Myeloma Cells via Mcl‐1 Downregulation

Inhibitors of cyclin‐dependent kinases 9 have been developed as potential anticancer drugs for the treatment of multiple myeloma. We have previously prepared a library of arylazo‐3,5‐diaminopyrazole inhibitors of CDKs. Here, we describe a novel member, AAP1742 (CDK9 inhibition with IC₅₀ = 0.28 μm ), that reduces the viability of multiple myeloma cell lines in low micromolar concentrations. Consistent with inhibition of CDK9, AAP1742 decreases the phosphorylation of RNA polymerase II and inhibits mRNA synthesis of anti‐apoptotic proteins Mcl‐1, Bcl‐2, and XIAP, followed by apoptosis in the RPMI‐8226 cell line in a dose‐ and a time‐dependent manner. These results are consistent with the biochemical profile of AAP1742 and further suggest cellular inhibition of CDK9 as a possible target for anticancer drugs.     

Efficient Closed Loop Simulation of Do-It-Yourself Artificial Pancreas Systems

Background:Numerical simulations, also referred to as in silico trials, are nowadays the first step toward approval of new artificial pancreas (AP) systems. One suitable tool to run such simulations is the UVA/Padova Type 1 Diabetes Metabolic Simulator (T1DMS). It was used by Toffanin et al. to provide data about safety and efficacy of AndroidAPS, one of the most wide-spread do-it-yourself AP systems. However, the setup suffered from slow simulation speed. The objective of this work is to speed up simulation by implementing the algorithm directly in MATLAB^®/Simulink^®.Method:Firstly, AndroidAPS is re-implemented in MATLAB^® and verified. Then, the function is incorporated into T1DMS. To evaluate the new setup, a scenario covering 2 days in real time is run for 30 virtual patients. The results are compared to those presented in the literature.Results:Unit tests and integration tests proved the equivalence of the new implementation and the original AndroidAPS code. Simulation of the scenario required approximately 15 minutes, corresponding to a speed-up factor of roughly 1000 with respect to real time. The results closely resemble those presented by Toffanin et al. Discrepancies were to be expected because a different virtual population was considered. Also, some parameters could not be extracted from and harmonized with the original setup.Conclusions:The new implementation facilitates extensive in silico trials of AndroidAPS due to the significant reduction of runtime. This provides a cheap and fast means to test new versions of the algorithm before they are shared with the community.     

Immunomodulatory and antimicrobial non-mulberry Antheraea mylitta silk fibroin accelerates in vitro fibroblast repair and regeneration by protecting oxidative stress

The antimicrobial nature of Antharaea mylitta silk-fibroin (SF) is reported but antioxidant potential and the immunomodulatory role towards the fibroblast cell repair process is not explored. Polyurethane is reported to have inflammatory potential by mononuclear cells directed cytokine release, which can guide fibroblast repair. Present study demonstrates the conjunctive effect of inflammatory PU/SF to regulate the favorable shift from pro-inflammatory to anti-inflammatory cytokine stimulation for accelerated fibroblast repair. Minimal inhibitory concentration of SF was determined against pathogenic strains and the effect of SF was investigated for fibroblast NIH3T3 cell adhesion. SF doses (8, 8.5, 9 mg mL⁻¹) were found to be greater than both the IC₅₀ of DPPH scavenging and the ED₅₀ for NIH3T3 proliferation. Anti-lipid peroxidase (ALP) activity of SF doses and citric acid-treated NIH3T3 cells were compared under hydrogen peroxide (H₂O₂) induced oxidative stress. 9 mg mL⁻¹ SF showed greater ALP activity than the citric acid standard. SF-driven protection to oxidative damage was measured by viable cell fraction in trypan blue dye exclusion assay where 9 mg mL⁻¹ SF showed the highest viability (p ≤ 0.05). 9 mg mL⁻¹ SF was blended with PU for scaffold (w/v = 2 : 5, 2 : 7, 2 : 9) fabrication. The protective effect of PU/SF (2 : 5, 2 : 7, 2 : 9) against oxidative stress was verified by damaged cell survival in MTT assay and DNA quantification. The highest number of cells survived on PU/SF (2 : 9) at all intervals (p ≤ 0.01) upon oxidative damage; PU/SF (2 : 9) was also fabricated by employing the immobilization technique. Immobilized PU/SF (2 : 9) exhibited a greater zone of microbial inhibition, a higher extent of inhibition to microbial adherence, and caused more LDH release from bacterial cell membrane due to membrane rupture, resulting in bacterial cell death (E. coli, K. pneumoniae, P. aeruginosa, S. aureus) compared to the experimental results shown by blended PU/SF (2 : 9). The protective nature of PU/SF (2 : 9) against oxidative stress was ensured through the LDH activity of damaged NIH3T3 cells. Initial raised IL-6, TNF-alpha (pro-inflammatory cytokines) and lowered IL-8, IL-10 (anti-inflammatory cytokine) profiles coupled with fallen IL-6, TNF-alpha, and elevated IL-8, IL-10 at later hours synergistically progress the inflammatory phase of in vitro scratch wound repair in mononuclear culture treated by PU/SF (2 : 9).

Initially SF accelerated pro-inflammatory cytokines, restricted anti-inflammatory cytokines; later it regulated in reverse order. SF potentially eradicated ROS and promoted Ki-67 cellular regeneration whereas pristine PU could not.     

Liver is widely eaten by preschool children in the Northern Cape province of South Africa: Implications for routine vitamin A supplementation

Previous research has demonstrated a virtual absence of vitamin A deficiency and adequacy of vitamin A intake through consumption of liver in preschool children of a community in the Northern Cape province of South Africa where sheep farming is common, and liver, an exceptionally rich source of vitamin A, is frequently eaten. Only 60–75 g of liver per month is needed to meet the vitamin A requirement of preschool children. Because this may have implications for routine vitamin A supplementation, and because liver consumption for the rest of the province is unknown, the study aim was to establish the prevalence and frequency of liver intake in a provincial‐wide survey. An unquantified liver‐specific food frequency questionnaire, covering a period of 1 month, complemented by a 1‐year recall, was administered to mothers of 2‐ to 5‐year‐old children (n = 2,864) attending primary health care facilities in all five districts and 26 subdistricts. A total of 86% of children were reported to eat liver, which was eaten in all districts by at least 80% of children. The overall median frequency of liver intake was 1.0 [25th, 75th percentiles: 0.5, 3.0] times per month and ranged from 1.0 [0.3, 2.0] to 2.0 [1.0, 4.0] for the various districts. Based on a previously reported portion size of 66 g, these results suggest vitamin A dietary adequacy in all districts and possibly also vitamin A intake exceeding the Tolerable Upper Intake Level in some children. Routine vitamin A supplementation in this province may not be necessary and should be reconsidered.     

Singleton exome sequencing of 90 fetuses with ultrasound anomalies revealing novel disease-causing variants and genotype–phenotype correlations

Exome sequencing has been increasingly implemented in prenatal genetic testing for fetuses with morphological abnormalities but normal rapid aneuploidy detection and microarray analysis. We present a retrospective study of 90 fetuses with different abnormal ultrasound findings, in which we employed the singleton exome sequencing (sES; 75 fetuses) or to a lesser extent (15 fetuses) a multigene panel analysis of 6713 genes as a primary tool for the detection of monogenic diseases. The detection rate of pathogenic or likely pathogenic variants in this study was 34.4%. The highest diagnostic rate of 56% was in fetuses with multiple anomalies, followed by cases with skeletal or renal abnormalities (diagnostic rate of 50%, respectively). We report 20 novel disease-causing variants in different known disease-associated genes and new genotype–phenotype associations for the genes KMT2D, MN1, CDK10, and EXOC3L2. Based on our data, we postulate that sES of fetal index cases with a concurrent sampling of parental probes for targeted testing of the origin of detected fetal variants could be a suitable tool to obtain reliable and rapid prenatal results, particularly in situations where a trio analysis is not possible.Subject terms: Genetics research, Disease genetics     

The Effect of Mask Style and Fabric Selection on the Comfort Properties of Fabric Masks

The purpose of fabric masks in the prevention of the spread of COVID-19 often requires that the masks be worn for extended periods without removal. The management of the conditions in the micro-climate inside the masks is important to keep the wearer comfortable and enhance user compliance. In this study, the effect of mask design and fabric type on the micro-climate was investigated using thermocron iButtons to record the temperature and humidity inside the masks. It was found that the mask style, and its effect on the amount of air incorporated in the micro-climate, had a significant influence on the factors that determine the temperature and humidity levels. In the shaped masks, the impact of the mask design on the results was stronger than the effect of fabric type. In the folded masks that fit snugly around the face, the effect of fabric type was significant, and both fibre composition and fabric structure contributed to the differences in the performance of the three fabrics tested. In the case of the masks with an inserted filter, a significant amount of trapped still air in the fabric layers and the increased mask stiffness had the strongest effect on the temperature and humidity inside the masks. Significant differences were also found in the temperatures recorded in the different time segments, highlighting the importance of conducting comfort evaluations over a long enough time to prevent false interpretations. The results of this study emphasize the importance of considering all the components of mask design, namely style, fibre type, and fabric structure, in the development of masks to enhance user compliance.