Microfluidics-assisted rapid generation of tubular cell-laden microgel inside glass capillaries

Drug screening using engineered blood vessels (EBVs) faces considerable barriers in approximating the conditions of an in vivo environment. To address this issue, we have introduced a microfluidic system for cell-laden tubular microgels. N-Carboxyethyl chitosan crosslinked with oxidized dextran was used for in situ gelable tubular scaffolds. The microfluidic system consisted of four glass capillaries that generated a coaxial flow of pre-polymer and phosphate buffered solutions. It rapidly produced cell-laden tubular microgels inside glass capillaries. The mechanical strength of the tubular microgels was suitable for their application as EBVs, with a maximum Young’s modulus of 12.2 ± 1.9 kPa. In vitro cell studies using human umbilical vein endothelial cells verified the biocompatibility and non-cytotoxicity of the gelation and fabrication process. Thus, in situ gelable cell-laden tubular microgels can be a potential platform for screening drugs to treat blood vessel diseases.     

Antibacterial activity of silver nanoparticle-coated fabric and leather against odor and skin infection causing bacteria

We present a simple, eco-friendly synthesis of silver and gold nanoparticles using a natural polymer pine gum solution as the reducing and capping agent. The pine gum solution was combined with silver nitrate (AgNO₃) or a chloroauric acid (HAuCl₄) solution to produce silver nanoparticles (AgNPs) and gold nanoparticles (AuNPs), respectively. The reaction process was simple; formation of the nanoparticles was achieved by autoclaving the silver and gold ions with the pine gum. UV–Vis spectra showed surface plasmon resonance (SPR) for silver and gold nanoparticles at 432 and 539 nm, respectively. The elemental forms of AgNPs and AuNPs were confirmed by energy-dispersive X-ray spectroscopy (EDX). Fourier transform infrared spectroscopy (FTIR) showed the biomolecules present in the pine gum, AgNPs, and AuNPs. Transmission electron microscopy (TEM) images showed the shape and size of AgNPs and AuNPs. The crystalline nature of synthesized AgNPs and AuNPs was confirmed by X-ray crystallography [X-ray diffraction (XRD)]. Application of synthesized AgNPs onto cotton fabrics and leather, in order to evaluate their antibacterial properties against odor- or skin infection-causing bacteria, is also discussed. Among the four tested bacteria, AgNP-coated cotton fabric and leather samples displayed excellent antibacterial activity against Brevibacterium linens.     

Arabidopsis Fused kinase TWO-IN-ONE dominantly inhibits male meiotic cytokinesis

Arabidopsis Fused kinase TWO-IN-ONE (TIO) controls phragmoplast expansion through its interaction with the Kinesin-12 subfamily proteins that anchor the plus ends of interdigitating microtubules in the phragmoplast midzone. Previous analyses of loss-of-function mutants and RNA interference lines revealed that TIO positively controls both somatic and gametophytic cell cytokinesis; however, knowledge of the full spectrum of TIO functions during plant development remains incomplete. To characterize TIO functions further, we expressed TIO and a range of TIO variants under control of the TIO promoter in wild-type Arabidopsis plants. We discovered that TIO-overexpressing transgenic lines produce enlarged pollen grains, arising from incomplete cytokinesis during male meiosis, and show sporophytic abnormalities indicative of polyploidy. These phenotypes arose independently in TIO variants in which either gametophytic function or the ability of TIO to interact with Kinesin-12 subfamily proteins was abolished. Interaction assays in yeast showed TIO to bind to the AtNACK2/TETRASPORE, and plants doubly homozygous for kinesin-12a and kinesin-12b knockout mutations to produce enlarged pollen grains. Our results show TIO to dominantly inhibit male meiotic cytokinesis in a dosage-dependent manner that may involve direct binding to a component of the canonical NACK-PQR cytokinesis signaling pathway.     

A robust,simple, high‐throughput technique for time‐resolved plant volatile analysis in field experiments

Plant volatiles (PVs) mediate interactions between plants and arthropods, microbes and other plants, and are involved in responses to abiotic stress. PV emissions are therefore influenced by many environmental factors, including herbivore damage, microbial invasion, and cues from neighboring plants, and also light regime, temperature, humidity and nutrient availability. Thus, an understanding of the physiological and ecological functions of PVs must be based on measurements reflecting PV emissions under natural conditions. However, PVs are usually sampled in the artificial environments of laboratories or climate chambers. Sampling of PVs in natural environments is difficult, being limited by the need to transport, maintain and provide power to instruments, or use expensive sorbent devices in replicate. Ideally, PVs should be measured in natural settings with high replication, spatio‐temporal resolution and sensitivity, and modest costs. Polydimethylsiloxane (PDMS), a sorbent commonly used for PV sampling, is available as silicone tubing for as little as 0.60 € m^?1 (versus 100–550 € each for standard PDMS sorbent devices). Small pieces of silicone tubing (STs) of various lengths from millimeters to centimeters may be added to any experimental setting and used for headspace sampling, with little manipulation of the organism or headspace. STs have sufficiently fast absorption kinetics and large capacity to sample plant headspaces over a timescale of minutes to hours, and thus can produce biologically meaningful ‘snapshots’ of PV blends. When combined with thermal desorption coupled to GC–MS (a 40‐year‐old widely available technology), use of STs yields reproducible, sensitive, spatio‐temporally resolved quantitative data from headspace samples taken in natural environments.     

External Validation and Evaluation of Reliability and Validity of the S-ReSC Scoring System to Predict Stone-Free Status after Percutaneous Nephrolithotomy

Objectives

The Seoul National University Renal Stone Complexity (S-ReSC) scoring system was developed to predict the stone-free rate (SFR) after single-tract percutaneous nephrolithotomy (PCNL). This study is an external validation of this scoring system.

Materials and methods

A retrospective review included 327 patients who underwent PCNL at 2 tertiary referral centers. The S-ReSC score was assigned from 1 to 9 based on the number of sites involved. The stone free status was defined as either complete clearance or clinically insignificant residual fragments <4 mm in size at 1 month follow-up imaging. Inter-observer and test-retest reliabilities were evaluated. The statistical performance of the prediction model was assessed by its predictive accuracy, predictive probability, and clinical usefulness.

Results

The overall SFR was 65.4%. SFRs were 83.9%, 47.6%, and 21.4% in low (1–2), intermediate (3–4), and high (5–9) score groups, respectively, with significant differences (P<0.001). Inter-observer and test-retest reliabilities revealed almost perfect agreements. External validation of the S-ReSC scoring system revealed an AUC of 0.731 (95% CI 0.675–0.788). The AUC of 3-titered S-ReSC score groups was 0.691 (95% CI, 0.629–0.753). The calibration plot showed that the predicted probability of SFR had a concordance comparable to that of the observed frequency. The Hosmer–Lemeshow goodness-of-fit statistic revealed an adequate performance of the predictive model (P = 0.10). Inter-observer and test-retest reliability showed a good level of agreement.

Conclusions

The S-ReSC scoring system is useful in predicting the post-PCNL SFR and in describing the complexity of renal stones.     

Detection of Retinitis Pigmentosa by Differential Interference Contrast Microscopy

Differential interference contrast microscopy is designed to image unstained and transparent specimens by enhancing the contrast resulting from the Nomarski prism-effected optical path difference. Retinitis pigmentosa, one of the most common inherited retinal diseases, is characterized by progressive loss of photoreceptors. In this study, Differential interference contrast microscopy was evaluated as a new and simple application for observation of the retinal photoreceptor layer and retinitis pigmentosa diagnostics and monitoring. Retinal tissues of Royal College of Surgeons rats and retinal-degeneration mice, both well-established animal models for the disease, were prepared as flatmounts and histological sections representing different elapsed times since the occurrence of the disease. Under the microscopy, the retinal flatmounts showed that the mosaic pattern of the photoreceptor layer was irregular and partly collapsed at the early stage of retinitis pigmentosa, and, by the advanced stage, amorphous. The histological sections, similarly, showed thinning of the photoreceptor layer at the early stage and loss of the outer nuclear layer by the advanced stage. To count and compare the number of photoreceptors in the normal and early-retinitis pigmentosa-stage tissues, an automated cell-counting program designed with MATLAB, a numerical computing language, using a morphological reconstruction method, was applied to the differential interference contrast microscopic images. The number of cells significantly decreased, on average, from 282 to 143 cells for the Royal College of Surgeons rats and from 255 to 170 for the retinal-degeneration mouse. We successfully demonstrated the potential of the differential interference contrast microscopy technique’s application to the diagnosis and monitoring of RP.