Washed microbiota transplantation for Crohn's disease: A metagenomic, metatranscriptomic, and metabolomic-based study.

BACKGROUND: Fecal microbiota transplantation (FMT) is a promising therapeutic approach for treating Crohn's disease (CD). The new method of FMT, based on the automatic washing process, was named as washed microbiota transplantation (WMT). Most existing studies have focused on observing the clinical phenomena. However, the mechanism of action of FMT for the effective management of CD-particularly in-depth multi-omics analysis involving the metagenome, metatranscriptome, and metabolome-has not yet been reported. AIM: To assess the efficacy of WMT for CD and explore alterations in the microbiome and metabolome in response to WMT. METHODS: We conducted a prospective, open-label, single-center clinical study. Eleven CD patients underwent WMT. Their clinical responses (defined as a decrease in their CD Activity Index score of > 100 points) and their microbiome (metagenome, metatranscriptome) and metabolome profiles were evaluated three months after the procedure. RESULTS: Seven of the 11 patients (63.6%) showed an optimal clinical response three months post-WMT. Gut microbiome diversity significantly increased after WMT, consistent with improved clinical symptoms. Comparison of the metagenome and metatranscriptome analyses revealed consistent alterations in certain strains, such as Faecalibacterium prausnitzii, Roseburia intestinalis, and Escherichia coli. In addition, metabolomics analyses demonstrated that CD patients had elevated levels of various amino acids before treatment compared to the donors. However, levels of vital amino acids that may be associated with disease progression (e.g., L-glutamic acid, gamma-glutamyl-leucine, and prolyl-glutamine) were reduced after WMT. CONCLUSION: WMT demonstrated therapeutic efficacy in CD treatment, likely due to the effective reconstruction of the patient's microbiome. Multi-omics techniques can effectively help decipher the potential mechanisms of WMT in treating CD.

Production of Trans-Cinnamic Acid by Immobilization of the Bambusa oldhamii BoPAL1 and BoPAL2 Phenylalanine Ammonia-Lyases on Electrospun Nanofibers.

Phenylalanine ammonia-lyase (PAL) catalyzes the nonoxidative deamination of phenylalanine to yield trans-cinnamic acid and ammonia. Recombinant Bambusa oldhamii BoPAL1/2 proteins were immobilized onto electrospun nanofibers by dextran polyaldehyde as a cross-linking agent. A central composite design (CCD)-response surface methodology (RSM) was utilized to optimize the electrospinning parameters. Escherichia coli expressed eBoPAL2 exhibited the highest catalytic efficiency among four enzymes. The optimum conditions for fabricating nanofibers were determined as follows: flow rate of 0.10 mL/h, voltage of 13.8 kV, and distance of 13 cm. The response surface models were used to obtain the smaller the fiber diameters as well as the highest PAL activity in the enzyme immobilization. Compared with free BoPALs, immobilized BoPALs can be reused for at least 6 consecutive cycles. The remained activity of the immobilized BoPAL proteins after storage at 4 °C for 30 days were between 75 and 83%. In addition, the tolerance against denaturants of the immobilized BoPAL proteins were significantly enhanced. As a result, the dextran polyaldehyde natural cross-linking agent can effectively replace traditional chemical cross-linking agents for the immobilization of the BoPAL enzymes. The PAL/nylon 6/polyvinyl alcohol (PVA)/chitosan (CS) nanofibers made are extremely stable and are practical for industrial applications in the future.

NLIP and HAD-like Domains of Pah1 and Lipin 1 Phosphatidate Phosphatases Are Essential for Their Catalytic Activities.

Saccharomyces cerevisiae Pah1 phosphatidate phosphatase (PAP) catalyzes the dephosphorylation of phosphatidate to yield diacylglycerol, controlling phospholipids and triacylglycerol metabolisms. Pah1 and human Lipin 1 are intrinsically disordered proteins with 56% and 43% unfolded regions, respectively. Truncation analysis of the conserved and non-conserved regions showed that N- and C-conserved regions are essential for the catalytic activity of Pah1. PAP activities can be detected in the conserved N-terminal Lipin (NLIP) domain and C-terminal Lipin (CLIP)/haloacid dehalogenase (HAD)-like domain of Pah1 and Lipin 1, suggesting that the evolutionarily conserved domains are essential for the catalytic activity. The removal of disordered hydrophilic regions drastically reduced the protein solubility of Pah1. Thioredoxin is an efficient fusion protein for production of soluble NLIP-HAD recombinant proteins in Escherichia coli.

[Screening of Amendments for Simultaneous Cd and As Immobilization in Soil].

Simultaneously reducing the availability of Cd and As is difficult owing to converse chemical behaviors of Cd and As in soil. In this study, amendments that can simultaneously immobilize Cd and As in soil were determined by an pure soil culture experiment in which flooding and wetting were performed for 30 d each. The effects of sepiolite (Sep), modified sepiolite (IMS and Sep-FM), steel slag (SS), and iron modified biochar (Fe-Bio) on soil pH, Eh, Cd, and As concentrations in pore water, and Cd and As fractions in soil were investigated. It showed that Sep (1%, 2.5%), IMS (1%, 2.5%), Sep-FM (1%, 2.5%), and SS (1%, 5%) treatments increased soil pH value and decreased Eh value and Cd concentrations in soil solution. In addition, As concentrations in soil solution treated with high doses of IMS (2.5%) and SS (5%) were lower than that of CK treatment during the whole incubation period. However, Fe-bio treatment decreased soil pH and increased Eh value and only decreased Cd and As concentrations in soil solution under wet conditions. Compared with the control, the application of the above amendments promoted the transformation of Cd fraction from exchangeable to reducible, oxidizable, and residual. High application rates of IMS (2.5%), Sep-FM (2.5%), and SS (5%) also reduced available As fraction (non-specifically sorbed and specifically-sorbed As fraction), and increased amorphous and poorly-crystalline hydrated Fe and Al oxide-bound As. On the contrary, Fe-bio treatment increased the fractions of non-specifically sorbed, specifically sorbed and residual As in soil. In short, IMS, Sep-FM, and SS are potential materials for remediation of Cd and As contaminated soil. They can effectively immobilize soil Cd and As and promote their transformation to the fractions that plants are difficult to uptake.

Using Convolutional Neural Networks to Measure the Physiological Age of Caenorhabditis elegans.

Caenorhabditis elegans (C. elegans) is a popular and excellent model for studies of aging due to its short lifespan. Methods for precisely measuring the physiological age of C. elegans are critically needed, especially for antiaging drug screening and genetic screening studies. The effects of various antiaging interventions on the rate of aging in the early stage of the aging process can be determined based on the quantification of physiological age. However, in general, the age of C. elegans is evaluated via human visual inspection of morphological changes based on personal experience and subjective judgment. For example, the rate of motor activity decay has been used to predict lifespan in early- to mid-stage aging. Using image processing, the physiological age of C. elegans can be measured and then classified into periods or classes from childhood to elderhood (e.g., 3 periods comprising days 0-2, 4-6 and 10-12) by using texture entropy (Shamir, L. et al., 2009). Our dataset consists of 913 microscopic images of C. elegans, with approximately 60 images per day from day 1 to day 14 of adulthood. We present quantitative methods to measure the physiological age of C. elegans with convolution neural networks (CNNs), which can measure age with a granularity of days rather than periods. The methods achieved a mean absolute error (MAE) of less than 1 day for the measured age of C. elegans. In our experiments, we found that after training and testing our dataset, 5 popular CNN models, 50-layer residual network (ResNet50), InceptionV3, InceptionResNetV2, 16-layer Visual Geometry Group network (VGG16) and MobileNet, measured the physiological age of C. elegans with an average testing MAE of 1.58 days. Furthermore, based on the results, we propose two models, one model for linear regression analysis and the other model for logistic regression, that combine a CNN model and a new attribute: curved_or_straight. The linear regression analysis model achieved a test MAE of 0.94 days; the logistic regression model achieved an accuracy of 84.78 percent with an error tolerance of 1 day.

Recycling furnace slag and fly ash from industrial byproducts to produce slag/ash based zeolite as a new adsorbent material.

This study recycling the industrial byproducts of furnace slag and fly ash to produce slag/ash based zeolite. A scanning electron microscopic/energy dispersive X-ray spectroscopic (SEM/EDS) analysis of this zeolite indicates a high SiO2 content of 53.94%, an Al2O3 content of 23.20%, a silicon-to-aluminum ratio of 2.049, a density of 2.88 g/cm3, and a water content of 0.13% while the zeolite appears as a porous crystalline structure. Results of weighing experiments revealed effective adsorption of liquid salad oil and highly volatile organic solvents after reusing the zeolite up to 6 times. When an adsorbed liquid pollutant was desorbed and the heating temperature exceeded 170 °C and 350 °C, the samples exhibited two exothermic reactions, respectively, the mean maximum enthalpy were 0.427 and 0.461 mW, and the mean per gram of accumulated heat were 80.92 and 45.64 J/g. For all samples, the mean maximum loss of mass rate was 9.26%. Analogously, for gaseous pollutants, the samples exhibited an exothermic reaction when the heating temperature exceeded 180 °C; the average maximum enthalpy was 0.395 mW, the average per gram of accumulated heat was 119.60 J/g, and the average maximum loss of mass rate was 4.79%. This slag/ash based zeolite has the advantages of low cost, high thermal stability, reusability, etc., and can be used as a new adsorbent material for indoor ventilation equipment.

Hydrogenation of High Molecular Weight Bisphenol A Type Epoxy Resin BE503 in a Functional and Greener Solvent Mixture Using a Rh Catalyst Supported on Carbon Black.

A functional greener solvent mixture containing water, isopropyl alcohol (IPA) and ethyl acetate with the ratio 10:20:70 (wt%) was found to accelerate hydrogenation of bisphenol A type epoxy resin BE503 with a molecular weight of 1500 through an on-water mechanism, and led to an increased H2 availability, due to high solubility of H2 in IPA. Different carbon-based supports were tested and VulcanXC72 was found as the best support among the tested carbon-based ones as it possessed the highest amount of electron deficient promoter, RhOx. The catalyst, Rh5/VulcanXC72-polyol, synthesized by the microwave assisted polyol method, yielded a 100% hydrogenation of aromatic rings with an epoxy ring opening below 20.0% at 50 °C and a H2 pressure of 1000 psi in 2.25 h. Intrinsic activation energies for the hydrogenation of aromatic rings and epoxy ring opening were experimentally estimated and a mechanism for the hydrogenation of BE503 was proposed, wherein the hydrogenation of aromatic rings and epoxy ring opening in BE503 proceeded simultaneously in parallel and in-series with parallel being the major pathway.

An edge-lit volume holographic optical element for an objective turret in a lensless digital holographic microscope.

In this paper, we propose and demonstrate the use of an edge-lit volume holographic optical element (EL-VHOE) as a reference waveguide to reduce the volume of a lensless digital holographic microscope. Additionally, a hybrid lensless Fourier transform digital holography is applied to make the EL-VHOE function as an objective turret. It used a spherical wave in the object beam of the EL-VHOE, which served as the reference beam of the microscope. Another sheared spherical wave was used to illuminate the sample. The longitudinal position of the spherical reference beam is changeable. It was shown that the tradeoff between resolution and field of view can be adjusted by changing the longitudinal position of the spherical reference beam. The corresponding experimental results matched the simulational and theoretical predictions. A resolution of approximately 3.11 µm was achieved when the object distance was 6 mm and the longitudinal distance of the spherical reference was 10 mm.

Automatic Sleep Staging in Patients With Obstructive Sleep Apnea Using Single-Channel Frontal EEG.

STUDY OBJECTIVES: Reliable sleep staging is difficult to obtain from home sleep testing for diagnosis of obstructive sleep apnea (OSA), especially when it is self-applied. Hence, the current study aimed to develop a single frontal electroencephalography-based automatic sleep staging system (ASSS). METHODS: The ASSS system was developed on a clinical dataset, with a high percentage of participants with OSA. The F4-M1 signal extracted from 62 participants (62.9% having OSA) was used to build a four-stage classifier. Performance of the ASSS was tested in a holdout set of 58 patients (60.3% having OSA) with epoch-by-epoch and whole-night agreement for sleep staging compared with expert scoring of polysomnography. RESULTS: Mean all-stage percentage agreement was 75.52% (95% confidence interval, 72.90 to 78.13) (kappa 0.62; 95% confidence interval, 0.58 to 0.65), with mean percentage agreement for wake, light sleep, deep sleep (DS), and rapid eye movement of 78.04%, 70.97%, 83.65%, and 75.00%, respectively. The whole-night agreement was good-excellent (intraclass correlation coefficient, 0.74 to 0.88) for sleep onset latency, wake after sleep onset, total sleep time, and sleep efficiency. Compared to the non-OSA subset, the OSA subset had lower agreement for DS. CONCLUSIONS: Our results indicate that a single-channel F4-M1 based ASSS was sufficient for sleep staging in a population with a high percentage of participants with OSA.

Reversible Self-Assembly Nanovesicle of UCST Response Prepared with Multi-l-arginyl-poly-l-aspartate Conjugated with Polyethylene Glycol.

Multi-L-arginyl-poly-L-aspartate (MAPA), also known as cyanophycin, containing a backbone of polyaspartate with arginine and lysine as side chains, was prepared with recombinant Escherichia coli. The insoluble part (iMAPA) was conjugated with polyethylene glycol (PEG) at two different levels, high (iMAPA(PEG)h) and low (iMAPA(PEG)l). Both levels of conjugation exhibited UCST (upper critical solution temperature)-type responses in the pH range of 3-10 at a concentration of 2 mg/mL. The cloud-point temperature of each conjugate also showed a positive correlation with concentration in PBS, falling between 20 to 58 °C at a concentration from 0.1 to 3 mg/mL. Hysteresis was observed to follow approximate paths under the same condition during repeated heating and cooling. Notably, the reversible formation of core-shell vesicles appeared at room temperature in PBS with a size of around 25 to 60 nm, as measured by DLS and observed under TEM. The reversibility was further employed to encapsulate doxorubicin (Dox) at different weight ratios of Dox to iMAPA(PEG)h. An encapsulation efficiency could reach as high as 70% with an equivalent loading capacity of 1.5 mg Dox/mg iMAPA(PEG)h. The Dox-loaded vesicles stayed stable at 4 °C for up to 4 weeks, with a minimal leakage below 2% and a slightly dilated morphology. Temperature-triggered release of Dox from the vesicles could be achieved by a step change of 5 °C successively from 37 to 62 °C in an effort to induce an initial 10% release at 37 °C gradually to complete release at 62 °C.

Green and rapid synthesis of zirconium metal-organic frameworks via mechanochemistry: UiO-66 analog nanocrystals obtained in one hundred seconds.

A UiO-66 analog was synthesized in 100 s using water-assisted grinding. The linker solubility suggested that tetrafluorobenzene-1,4-dicarboxylic acid was the best linker. Zr-metal-organic framework nanocrystals displayed good topologies and hydrophobicities, and high water/thermal stabilities. The less amorphous complex led to higher porosities and pore volumes with a 60 min grinding time.

Bone formation at recombinant human bone morphogenetic protein-2-coated titanium implants in the posterior maxilla (Type IV bone) in non-human primates.

BACKGROUND: Studies using ectopic rodent and orthotopic canine models (Type II bone) have shown that titanium porous oxide (TPO) surface implants adsorbed with recombinant human bone morphogenetic protein-2 (rhBMP-2) induce local bone formation including osseointegration. The objective of this study was to evaluate local bone formation and osseointegration at such implants placed into Type IV bone. MATERIAL AND METHODS: rhBMP-2-coated implants were installed into the edentulated posterior maxilla in eight young adult Cynomolgus monkeys: four animals each received three TPO implants adsorbed with rhBMP-2 (2.0 mg/ml) and four animals each received three TPO implants adsorbed with rhBMP-2 (0.2 mg/ml). Contra-lateral jaw quadrants received three TPO implants without rhBMP-2 (control). Treatments were alternated between left and right jaw quadrants. Mucosal flaps were advanced and sutured to submerge the implants. The animals received fluorescent bone markers at weeks 2, 3, 4, and at week 16 when they were euthanized for histologic analysis. RESULTS: Clinical healing was uneventful. Extensive local bone formation was observed in animals receiving implants adsorbed with rhBMP-2 (2.0 mg/ml). The newly formed bone exhibited a specific pinpoint bone-implant contact pattern regardless of rhBMP-2 concentration resulting in significant osseointegration; rhBMP-2 (2.0 mg/ml): 43% and rhBMP-2 (0.2 mg/ml): 37%. Control implants exhibited a thin layer of bone covering a relatively larger portion of the implant threads. Thus, TPO control implants bone exhibited significantly greater bone-implant contact ( approximately 75%; p<0.05). There were no statistically significant differences between rhBMP-2-coated and control implants relative to any other parameter including peri-implant and intra-thread bone density. CONCLUSION: rhBMP-2-coated TPO implants enhanced/accelerated local bone formation in Type IV bone in a dose-dependent fashion in non-human primates resulting in significant osseointegration. rhBMP-2-induced de novo bone formation did not reach the level of osseointegration observed in native resident bone within the 16-week interval.

Bone morphogenetic proteins: a realistic alternative to bone grafting for alveolar reconstruction.

Preclinical studies have shown that rhBMP-2 induces normal physiologic bone in clinically relevant defects in the craniofacial skeleton. The newly formed bone assumes characteristics of the adjacent resident bone and allows placement, osseointegration/re-osseointegration, and functional loading of endosseous implants. Clinical studies optimizing dose, delivery technologies, and conditions for stimulation of bone growth will bring about a new era in dentistry. The ability to predictably promote osteogenesis through the use of bone morphogenetic protein technologies is not far from becoming a clinical reality and will have an astounding effect on how dentistry is practiced.

Bone formation at titanium porous oxide (TiUnite) oral implants in type IV bone.

BACKGROUND: Several oral implant design advances have been suggested to overcome poor bone quality, an impediment for successful implant treatment. A novel titanium porous oxide (TPO) surface has been shown to offer favorable results in several settings. The objective of this study was to evaluate the local bone formation and osseointegration at TPO-modified implants in type IV bone. METHOD: Three TPO surface-modified implants (TiUnite) were installed into the edentulated posterior maxilla in each of 8 Cynomolgus monkeys. The animals were injected with fluorescent bone labels at 2, 3, 4 and 16 weeks post-surgery and were euthanized at week 16 when block biopsies were collected for histologic analysis. RESULTS: The predominant observation of the TPO implant surface was a thin layer of new bone covering most of the implant threads. Mean (+/-SE) bone-implant contact for the whole study group was 74.1 +/- 4.8%. There was a significant variability in bone-implant contact between animals (P = 0.0003) and between sites of the same animal (P < 0.0001). The variance in bone-implant contact was 30% larger among sites of the same animal than between different animals (187.5 vs. 144.8, respectively). There was a small but significant difference in bone density immediately outside, compared to within the threaded area of the implants (37.1 +/- 3.2% vs. 32.1 +/- 3.2%, P < 0.0001). Bone density outside the implant threads was significantly correlated (beta = 0.682, P < 0.0001) with the bone density within the threaded area. Bone density within the threaded area was significantly correlated (beta = 0.493, P = 0.0002) with bone-implant contact, whereas bone density outside the implant threads did not have a significant effect (beta = 0.232, P = 0.1). CONCLUSIONS: The results suggest that the TPO surface possesses a considerable osteoconductive potential promoting a high level of implant osseointegration in type IV bone in the posterior maxilla.