Engineering Rhodosporidium toruloides for production of 3-hydroxypropionic acid from lignocellulosic hydrolysate.

Microbial production of valuable bioproducts is a promising route towards green and sustainable manufacturing. The oleaginous yeast, Rhodosporidium toruloides, has emerged as an attractive host for the production of biofuels and bioproducts from lignocellulosic hydrolysates. 3-hydroxypropionic acid (3HP) is an attractive platform molecule that can be used to produce a wide range of commodity chemicals. This study focuses on establishing and optimizing the production of 3HP in R. toruloides. As R. toruloides naturally has a high metabolic flux towards malonyl-CoA, we exploited this pathway to produce 3HP. Upon finding the yeast capable of catabolizing 3HP, we then implemented functional genomics and metabolomic analysis to identify the catabolic pathways. Deletion of a putative malonate semialdehyde dehydrogenase gene encoding an oxidative 3HP pathway was found to significantly reduce 3HP degradation. We further explored monocarboxylate transporters to promote 3HP transport and identified a novel 3HP transporter in Aspergillus pseudoterreus by RNA-seq and proteomics. Combining these engineering efforts with media optimization in a fed-batch fermentation resulted in 45.4 g/L 3HP production. This represents one of the highest 3HP titers reported in yeast from lignocellulosic feedstocks. This work establishes R. toruloides as a host for 3HP production from lignocellulosic hydrolysate at high titers, and paves the way for further strain and process optimization towards enabling industrial production of 3HP in the future.

Tumor-targeted delivery of paclitaxel using low density lipoprotein-mimetic solid lipid nanoparticles.

Water-insoluble anticancer drugs, including paclitaxel, present severe clinical side effects when administered to patients, primarily associated with the toxicity of reagents used to solubilize the drugs. In efforts to develop alternative formulations of water-insoluble anticancer drugs suitable for intravenous administration, we developed biocompatible anticancer therapeutic solid lipid nanoparticles (SLNs), mimicking the structure and composition of natural particles, low-density lipoproteins (LDLs), for tumor-targeted delivery of paclitaxel. These therapeutic nanoparticles contained water-insoluble paclitaxel in the core with tumor-targeting ligand covalently conjugated on the polyethylene glycol (PEG)-modified surface (targeted PtSLNs). In preclinical human cancer xenograft mouse model studies, the paclitaxel-containing tumor-targeting SLNs exhibited pronounced in vivo stability and enhanced biocompatibility. Furthermore, these SLNs had superior antitumor activity to in-class nanoparticular therapeutics in clinical use (Taxol and Genexol-PM) and yielded long-term complete responses. The in vivo targeted antitumor activities of the SLN formulations in a mouse tumor model suggest that LDL-mimetic SLN formulations can be utilized as a biocompatible, tumor-targeting platform for the delivery of various anticancer therapeutics.

Rescue bowel preparation: same day 2 L polyethylene glycol addition, not superior to bisacodyl addition 7 days later.

BACKGROUND: The optimal colon-cleansing method after failure of bowel preparation (BP) for colonoscopy has not been established. AIMS: We aimed to compare BP rescue methods after failed initial BP and to identify risk factors for rescue BP failure. METHODS: Eighty-five patients with BP failure after 4 L polyethylene glycol (PEG) ingestion were prospectively enrolled from March 2008 to March 2012. A second colonoscopy was performed either on the same day after ingestion of another 2 L PEG (group A) or 1 week later after ingestion of 4 L PEG plus 20 mg oral bisacodyl (group B). Differences between groups in terms of BP quality and risk factors for a poor BP on the second colonoscopy were investigated. RESULTS: Median patient age was 59 years, 45 were male (52.9 %), and 17 (20 %) had poor BP on the second colonoscopy. For group B, the multivariable-adjusted odds ratio (OR) for poor BP on the second colonoscopy relative to group A was 0.68 (95 % confidence interval [CI], 0.16-2.95). Adequately ingested PEG during the initial colonoscopy was associated with poor BP on the second colonoscopy (OR 4.05; 95 % CI 1.04-15.75). The two groups had similar patient discomfort rates during the second BP. CONCLUSIONS: The two groups did not differ in rescue BP failure rate. Initial BP failure after adequate consumption of 4 L PEG may be a risk factor for rescue BP failure. A stricter BP regimen should be considered for these patients.

The use of assistive devices and social engagement among older adults: heterogeneity by type of social engagement and gender.

Little is known about the relationship between the use of assistive devices for functional loss and social engagement. This study investigated whether wearing assistive devices (dentures and hearing aids) is associated with social engagement among older adults in South Korea. Potential heterogeneity by type of social engagement (informal social contact versus formal social activity) and gender was also examined. This study analyzed data from 3725 individuals aged 65 or older collected over the course of 6 waves of the Korean Longitudinal Study of Ageing (KLoSA) from 2008 to 2018 (2150 women and 1575 men). Individual fixed effects models were used to account for unobserved individual-level heterogeneity that could confound the relationship between wearing assistive devices and social engagement. The fixed effects estimates showed that wearing assistive devices is associated with a decrease in formal social activity (b = - 0.034 for dentures and b = - 0.077 for hearing aids), but not informal social contact. Gender-stratified fixed effects revealed that the association between wearing assistive devices and social engagement is statistically significant only for men. In men, wearing assistive devices is associated with a decrease in formal social activity (b = - 0.049 for denture and b = - 0.095 hearing aids). The findings of this study suggest that wearing assistive devices may reduce the formal social activity of older adults, particularly men. Policymakers should consider developing interventions to help older adults overcome the stigma associated with assistive devices, which can hinder their social integration.

Postoperative Long-Term Monitoring of Mechanical Characteristics in Reconstructed Soft Tissues Using Biocompatible, Immune-Tolerant, and Wireless Electronic Sutures.

Accurate postoperative assessment of varying mechanical properties is crucial for customizing patient-specific treatments and optimizing rehabilitation strategies following Achilles tendon (AT) rupture and reconstruction surgery. This study introduces a wireless, chip-less, and immune-tolerant in vivo strain-sensing suture designed to continuously monitor mechanical stiffness variations in the reconstructed AT throughout the healing process. This innovative sensing suture integrates a standard medical suturing thread with a wireless fiber strain-sensing system, which incorporates a fiber strain sensor and a double-layered inductive coil for wireless readout. The winding design of Au nanoparticle-based fiber electrodes and a hollow core contribute to the fiber strain sensor's high sensitivity (factor of 6.2 and 15.1 pF for revised sensitivity), negligible hysteresis, and durability over 10,000 stretching cycles. To ensure biocompatibility and immune tolerance during extended in vivo periods, an antibiofouling lubricant layer was applied to the sensing suture. Using this sensing system, we successfully monitored the strain responses of the reconstructed AT in an in vivo porcine model. This facilitated the postoperative assessment of mechanical stiffness variations through a well-established analytical model during the healing period.

Lung-Mimetic Hydrofoam Sealant to Treat Pulmonary Air Leak.

Pulmonary air leak is the most common complication of lung surgery, contributing to post-operative morbidity in up to 60% of patients; yet, there is no reliable treatment. Available surgical sealants do not match the demanding deformation mechanics of lung tissue; and therefore, fail to seal air leak. To address this therapeutic gap, a sealant with structural and mechanical similarity to subpleural lung is designed, developed, and systematically evaluated. This "lung-mimetic" sealant is a hydrofoam material that has alveolar-like porous ultrastructure, lung-like viscoelastic properties (adhesive, compressive, tensile), and lung extracellular matrix-derived signals (matrikines) to support tissue repair. In biocompatibility testing, the lung-mimetic sealant shows minimal cytotoxicity and immunogenicity in vitro. Human primary monocytes exposed to sealant matrikines in vitro upregulate key genes (MARCO, PDGFB, VEGF) known to correlate with pleural wound healing and tissue repair in vivo. In rat and swine models of pulmonary air leak, this lung-mimetic sealant rapidly seals air leak and restores baseline lung mechanics. Altogether, these data indicate that the lung-mimetic sealant can effectively seal pulmonary air leak and promote a favorable cellular response in vitro.

Polypropylene microplastics promote metastatic features in human breast cancer.

Microplastics (MPs) are now a global issue due to increased plastic production and use. Recently, various studies have been performed in response to the human health risk assessment. However, these studies have focused on spherical MPs, which have smooth edges and a spherical shape and account for less than 1% of MPs in nature. Unfortunately, studies on fragment-type MPs are very limited and remain in the initial stages. In this study, we studied the effect that 16.4 µm fragment type polypropylene (PP) MPs, which have an irregular shape and sharp edges and form naturally in the environment, had on breast cancer. The detrimental effects of PPMPs on breast cancer metastasis were examined. Here, 1.6 mg/ml of PPMP, which does not induce cytotoxicity in MDA-MB-231, was used, and at this concentration, PPMP did not induce morphological changes or cellular migrating in the MDA-MB-231 and MCF-7 cells. However, PPMP incubation for 24 hours in the MDA-MB-231 cells significantly altered the level of cell cycle-related transcripts in an RNA-seq analysis. When confirmed by qRT-PCR, the gene expression of TMBIM6, AP2M1, and PTP4A2 was increased, while the transcript level of FTH1 was decreased. Further, secretion of the pro-inflammatory cytokine IL-6 from cancer cells was elevated with the incubation of PPMP for 12 hours. These results suggest that PPMP enhances metastasis-related gene expression and cytokines in breast cancer cells, exacerbating breast cancer metastasis.

Synthesis of aromatic nitriles using nonmetallic cyano-group sources.

Aromatic nitriles are prepared efficiently through transition-metal-mediated cyanation of aryl (pseudo)halides with metallic cyano-group sources, such as CuCN, KCN, NaCN, Zn(CN)(2), TMSCN, or K(4) [Fe(CN)(6)]. However, this approach often suffers from drawbacks, such as the formation of stoichiometric amounts of metal waste, the poisoning of the metal catalysts, or the generation of toxic HCN gas. As a result, a range of "nonmetallic" organic cyano-group sources have been explored for the cyanation of aryl halides and arene C-H bonds. This Minireview summarizes types of nonmetallic cyano-group sources and their applications in the preparation of aryl nitriles.

The effect of comprehensive oral care program on oral health and quality of life in patients undergoing radiotherapy for head and neck cancer: A quasi-experimental case-control study.

OBJECTIVES: The purpose of this study is to investigate the effect of the comprehensive oral care program on oral health status and symptoms in head and neck cancer (HNC) patients undergoing radiotherapy. METHODS: This was a quasi-experimental study using a non-equivalent control group in non-synchronized design. All participants including control and experimental group were asked for the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire H&N35 (EORTC QLQ-H&N35) and given an oral health education 4 times at baseline, immediate postradiotherapy, 3 months after radiotherapy, and 6 months after radiotherapy. In each visit except for final, the experimental group was given fluoride varnish application and fluoride mouth rinsing solution for daily use. Oral health examination for dental caries, plaque score (PS), bleeding on probing (BOP), and salivary flow rate was performed in baseline and 6 months after radiotherapy. Statistical analyses were done by paired t-tests and mixed ANCOVA repeated-measures analysis. RESULTS: From November 1, 2013 to October 31, 2015, a total 61 patients undergoing radiotherapy for HNC cancer were enrolled (30 in control and 31 in experimental groups). Decrease in salivary flow rate was comparable between 2 groups. Dental caries increased in control group (P = .006); PS and BOP were decreased in experimental group (P < .001 and .004, respectively). Experimental group showed lower swallowing, speech problems, and less sexuality scores in EORTC QLQ-H&N35 than control group. CONCLUSION: We found improvement in oral health and the quality of life in HNC patients with comprehensive oral care intervention by dental professionals. Communicating and cooperating between the healthcare and dental professionals is needed to raise the quality of health care services for HNC patients receiving radiotherapy.

Comparison of the IPSA and HIPO algorithms for interstitial tongue high-dose-rate brachytherapy.

PURPOSE: This study aimed to compare the inverse planning simulated annealing (IPSA) stochastic algorithm with the hybrid inverse planning and optimization (HIPO) algorithm for interstitial tongue high-dose-rate (HDR) brachytherapy. METHODS: Twenty patients who received radiotherapy for tongue cancer using interstitial HDR brachytherapy were retrospectively selected for this study. Oncentra Brachy v. 4.3 was used for IPSA and HIPO planning. Four to eight fixed catheter configurations were determined according to the target shape. During the optimization process, predetermined constrain values were used for each IPSA and HIPO plan. The dosimetric parameters and dwell time were analyzed to evaluate the performances of the plans. RESULTS: The total dwell time using IPSA was 4 seconds longer than that of HIPO. The number of active positions per catheter for the IPSA plans were approximately 2.5 fewer than those of the HIPO plans. The dose-volumetric parameters related to the clinical target volume with IPSA were lower than those with HIPO. In terms of the dose-volumetric parameters related to normal tissue, HIPO tended to associate with slightly higher values than IPSA, without statistical significance. After GrO, the target coverages were satisfied to clinical goal for all patients. The total dwell times was approximately increased by 10%. CONCLUSIONS: The IPSA and HIPO dose optimization algorithms generate similar dosimetric results. In terms of the dwell time, HIPO appears to be more beneficial.

Low-power and low-drug-dose photodynamic chemotherapy via the breakdown of tumor-targeted micelles by reactive oxygen species.

Tumor-targeted delivery of anticancer agents using nanocarriers has been explored to increase the therapeutic index of cancer chemotherapy. However, only a few nanocarriers are clinically available because the physiological complexity often compromises their ability to target, penetrate, and control the release of drugs. Here, we report a method which dramatically increases in vivo therapeutic drug efficacy levels through the photodynamic degradation of tumor-targeted nanocarriers. Folate-decorated poly(ethylene glycol)-polythioketal micelles are prepared to encapsulate paclitaxel and porphyrins. Photo-excitation generates reactive oxygen species within the micelles to cleave the polythioketal backbone efficiently and facilitate drug release only at the illuminated tumor site. Intravenous injection of a murine xenograft model with a low dose of paclitaxel within the micelles, one-milligram drug per kg (mouse), corresponding to an amount less than that of Taxol by one order of magnitude, induces dramatic tumor regression without any acute systemic inflammation responses or organ toxicity under low-power irradiation (55 mW cm-2) at 650 nm.

A disposable microfluidic device with a reusable magnetophoretic functional substrate for isolation of circulating tumor cells.

We describe an assembly-disposable microfluidic device based on a silicone-coated release polymer thin film. It consists of a disposable polymeric superstrate and a reusable functional substrate and they are assembled simply using vacuum pressure. The disposable polymeric superstrate is manufactured by bonding a silicone-coated release polymer thin film and a microstructured polydimethylsiloxane (PDMS) replica, containing only a simple structured microchannel. The reusable functional substrate generates an intricate energy field that can penetrate the micrometer-thick polymer film into the microchannel and control microfluids. This is the first report to introduce a silicone-coated release polyethylene terephthalate (PET) thin film as a bonding layer on a microstructured PDMS replica. The bonding strength was ∼600 kPa, which is the strongest among bonding methods of PDMS and PET polymer. Additionally, accelerated tests for bond stability and leakage demonstrated that the silicone-coated release PET film can form a very robust bond with PDMS. To demonstrate the usefulness of the proposed assembly-disposable microfluidic device, a lateral magnetophoretic microseparator was developed in an assembly-disposable microfluidic device format and was evaluated for isolating circulating tumor cells (CTCs) from patients with breast cancer.

Volumetric accuracy of cone-beam computed tomography.

PURPOSE: This study was performed to investigate the influence of object shape and distance from the center of the image on the volumetric accuracy of cone-beam computed tomography (CBCT) scans, according to different parameters of tube voltage and current. MATERIALS AND METHODS: Four geometric objects (cylinder, cube, pyramid, and hexagon) with predefined dimensions were fabricated. The objects consisted of Teflon-perfluoroalkoxy embedded in a hydrocolloid matrix (Dupli-Coe-Loid TM; GC America Inc., Alsip, IL, USA), encased in an acrylic resin cylinder assembly. An Alphard Vega Dental CT system (Asahi Roentgen Ind. Co., Ltd, Kyoto, Japan) was used to acquire CBCT images. OnDemand 3D (CyberMed Inc., Seoul, Korea) software was used for object segmentation and image analysis. The accuracy was expressed by the volume error (VE). The VE was calculated under 3 different exposure settings. The measured volumes of the objects were compared to the true volumes for statistical analysis. RESULTS: The mean VE ranged from -4.47% to 2.35%. There was no significant relationship between an object's shape and the VE. A significant correlation was found between the distance of the object to the center of the image and the VE. Tube voltage affected the volume measurements and the VE, but tube current did not. CONCLUSION: The evaluated CBCT device provided satisfactory volume measurements. To assess volume measurements, it might be sufficient to use serial scans with a high resolution, but a low dose. This information may provide useful guidance for assessing volume measurements.

A high-efficiency microfluidic device for size-selective trapping and sorting.

We report the development of a simple poly(dimethylsiloxane) microfluidic device for high-efficiency trapping and sorting of micron-size particles. In this device, hydrodynamic fluid flow through the sieve-like microfluidic channel sequentially fills the trap positions with particles of the trap size, and particles smaller than the trap size pass through the sieve and are trapped by smaller traps downstream. By incorporating side channels alongside the main channel, we were able to decouple the fluidic flow in one stage from the flows in the other stages. This decoupling allows us to modularize each stage of the device regardless of the size of the entire device. In our demonstration experiment with the prototype, we showed that more than 85% of the polystyrene microspheres (of sizes 15 µm, 6 µm and 4 µm) were sorted in the correct segment of the device that targets their respective sizes. Moreover, this high-efficiency device was able to trap all microspheres which were introduced into the device. Finally, we tested the device's ability to trap and sort three different species of waterborne parasites (Entamoeba, Giardia, and Cryptosporidium) and obtained excellent sorting performance.

A needle penetrating the stomach cavity after acupuncture.

Although acupuncture is known as a safe procedure that is widely used in many countries, complications including infection, hemorrhage, hematoma, pneumothorax, nerve damage, and cardiac tamponade have been reported. A needle penetrating the stomach after acupuncture, however, is very rare. Here, we report the case of 47-year-old woman who experienced abdominal pain 2 days after receiving acupuncture. Upper gastrointestinal endoscopy identified an approximately 2.5-cm long needle in the posterior wall of the antrum. The needle was removed endoscopically using rat tooth forceps with no complications.

ZrO2 surface chemically coated with hyaluronic acid hydrogel loading GDF-5 for osteogenesis in dentistry.

The objective of this study was to modify zirconium dioxide (ZrO(2)) with photo-cured hyaluronic acid hydrogel (pcHAgel), and to subsequently evaluate the bone regeneration potential of the modified ZrO(2). In the present study, HA grafted onto a ZrO(2) substrate was investigated for its biocompatibility and other properties. We describe the positive influences of ZrO(2) surface-modified with pcHAgel (Zr-3) containing two different loads of growth and differentiation factor-5 (GDF-5) to aid new bone formation as compared to the same amount of BMP-2 (Zr-4-7). We characterized the Zr-3 for their surface morphology and chemical properties. Atomic force microscopy (AFM), scanning electron microscope (SEM), and X-ray photoelectron spectroscopy (XPS) showed that the pcHAgel was successfully grafted onto the ZrO(2) surface. The sustained release of GDF-5 and BMP-2 were observed to occur in the Zr-4-7. In vitro cell tests showed a higher level of MG63 cell proliferation and differentiation on Zr-4-7 than on Zr-3. The Zr-3 is a good biomaterial to deliver osteogenic differentiation factors such as BMP-2 and GDF-5, and GDF-5 can be useful as an effective alternative to aid new bone formation as compared to BMP-2.

Fabrication of 1-dimensional porous hydroxyapatite and evaluation of its osteoconductivity.

Porous HA ceramics with 1-dimensional pore channels were fabricated to obtain controllable microstructure. 1-dimensional porous HA was objected to find out the optimum condition of bone ingrowth and also to facilitate the observation of osteocondutive behavior in porous HA. The porous structure was formed by burnt-out of polymeric fibers and the size of pores was determined by the diameter of polymeric fibers. The porosity could be varied by the thickness of HA slurry coated on polymeric fiber and by the thickness of HA tapes inserted between fiber layers. As result, 1-dimensional porous HA ceramics of this study have the uniform interconnection size (50-500 microm) and the linearly open pore structure. The compressive strength of 1-dimensional porous HA was 6-10 MPa similar to that of human cancellous bone. On the in vivo test, oteon-like osteoconduction in pore channel of 1-dimensional porous HA was observed, like what had been found in cortical bones. This osteon-like new bone grew from the surface to the center of pore channels. The 1-dimensional porous HA ceramics prepared in this study were very useful as a model system to observe bone ingrowth in the porous HA implants.