Research progress on differentiation and regulation of plant chromoplasts.

Carotenoids, natural tetraterpenoids found abundantly in plants, contribute to the diverse colors of plant non-photosynthetic tissues and provide fragrance through their cleavage products, which also play crucial roles in plant growth and development. Understanding the synthesis, degradation, and storage pathways of carotenoids and identifying regulatory factors represents a significant strategy for enhancing plant quality. Chromoplasts serve as the primary plastids responsible for carotenoid accumulation, and their differentiation is linked to the levels of carotenoids, rendering them a subject of substantial research interest. The differentiation of chromoplasts involves alterations in plastid structure and protein import machinery. Additionally, this process is influenced by factors such as the ORANGE (OR) gene, Clp proteases, xanthophyll esterification, and environmental factors. This review shows the relationship between chromoplast and carotenoid accumulation by presenting recent advances in chromoplast structure, the differentiation process, and key regulatory factors, which can also provide a reference for rational exploitation of chromoplasts to enhance plant quality.

Optimum Parameters in Ultrasound Coherent Plane Wave Compounding for High LPWV Estimation: Validation on Phantom and Feasibility in 10 Subjects.

OBJECTIVES: The aim of this study is to determine the optimum and fine values of the number and transmission angles of tilted plane waves for coherent plane-wave compounding (CPWC)-based high local pulse wave velocity (LPWV) estimation. METHODS: A Verasonics system incorporating a linear array probe L14-5/38 with 128 elements and a pulsatile pump, CompuFlow1000, were used to acquire radio frequency data of 3, 5, 7, and 9 tilted plane wave sequences with angle intervals from 0° to 12° with a coarse interval increment step of 1°, and the angle intervals from 0° to 2° with a fine interval increment step of 0.25° from a carotid vessel phantom with the LPWV of 13.42 ± 0.90 m/s. The mean value, standard deviation, and coefficients of variation (CV) of the estimated LPWVs were calculated to quantitatively assess the performance of different configurations for CPWC-based LPWV estimation. Ten healthy human subjects of two age groups were recruited to assess the in vivo feasibility of the optimum parameter values. RESULTS: The CPWC technique with three plane waves (PRF of 12 kHz corresponding to a frame rate of 4000 Hz) with an interval of 0.75° had LPWVs of 13.52 ± 0.08 m/s with the lowest CV of 1.84% on the phantom, and 5.49 ± 1.46 m/s with the lowest CV of 12.35% on 10 subjects. CONCLUSIONS: The optimum parameters determined in this study show the best repeatability of the LPWV measurements with a vessel phantom and 10 healthy subjects, which support further studies on larger datasets for potential applications.

[Genetic analysis of two patients with a rare Ael subtype].

OBJECTIVE: To analyze the genetic sequences of two patients with a rare Ael blood subgroup. METHODS: Two female patients undergoing treatment respectively for adenomyoma of the uterus and gastritis at the Second Affiliated Hospital, Yuying Children's Hospital of Wenzhou Medical University in June 2019 and September 2020 were selected as the study subjects. Their Ael subtypes were identified with a saline tube agglutination assay and absorption-emission assay. Sequence of the ABO gene Ael subtypes was determined by the Sanger method. The impact of genetic variants on the structural stability of N-acetylgalactosaminyl transferase (GTA) was analyzed with PyMOL software by constructing a structure predicted model. RESULTS: Both patients were determined as Ael blood subgroup. Sequencing result of patient 1 was ABO*O.01.02/ABO*O.01.02, which has resulted in a p.Thr88Profs*31 amino acid substitution. The sequencing result of patient 2 was ABO*Ael.06/ABO*O.01.02, in which c.425C>T and c.467C>T variants in exon 7 have led to p.Met142Thr and p.Pro156Leu substitutions. Prediction of the protein model speculated that the p.Met142Thr not only can change the binding of GTA protein with water molecules, but also the local hydrogen bond network of GTA, which may lead to decreased enzymatic activity. By contrast, the p.Pro156Leu variant has trivial effect on the structural stability of GTA. CONCLUSION: The molecular structure of Ael subtypes can be diverse. The genotypes of the two patients have been respectively determined as ABO*O.01.02/ABO*O.01.02 with a G261 deletion and ABO*Ael.06/ABO*O.01.02.

Generation of rat forebrain tissues in mice.

Interspecies blastocyst complementation (IBC) provides a unique platform to study development and holds the potential to overcome worldwide organ shortages. Despite recent successes, brain tissue has not been achieved through IBC. Here, we developed an optimized IBC strategy based on C-CRISPR, which facilitated rapid screening of candidate genes and identified that Hesx1 deficiency supported the generation of rat forebrain tissue in mice via IBC. Xenogeneic rat forebrain tissues in adult mice were structurally and functionally intact. Cross-species comparative analyses revealed that rat forebrain tissues developed at the same pace as the mouse host but maintained rat-like transcriptome profiles. The chimeric rate of rat cells gradually decreased as development progressed, suggesting xenogeneic barriers during mid-to-late pre-natal development. Interspecies forebrain complementation opens the door for studying evolutionarily conserved and divergent mechanisms underlying brain development and cognitive function. The C-CRISPR-based IBC strategy holds great potential to broaden the study and application of interspecies organogenesis.

Reconstructive interpolation for pulse wave estimation to improve local PWV measurement of carotid artery.

Ultrasonic transit time (TT)-based local pulse wave velocity (PWV) measurement is defined as the distance between two beam positions on a segment of common carotid artery (CCA) divided by the TT in the pulse wave propagation. However, the arterial wall motions (AWMs) estimated from ultrasonic radio frequency (RF) signals with a limited number of frames using the motion tracking are typically discrete. In this work, we develop a method involving motion tracking combined with reconstructive interpolation (MTRI) to reduce the quantification errors in the estimated PWs, and thereby improve the accuracy of the TT-based local PWV measurement for CCA. For each beam position, normalized cross-correlation functions (NCCFs) between the reference (the first frame) and comparison (the remaining frames) RF signals are calculated. Thereafter, the reconstructive interpolation is performed in the neighborhood of the NCCFs' peak to identify the interpolation-deduced peak locations, which are more exact than the original ones. According to which, the improved AWMs are obtained to calculate their TT along a segment of the CCA. Finally, the local PWV is measured by applying a linear regression fit to the time-distance result. In ultrasound simulations based on the pulse wave propagation models of young, middle-aged, and elderly groups, the MTRI method with different numbers of interpolated samples was used to estimate AWMs and local PWVs. Normalized root mean squared errors (NRMSEs) between the estimated and preset values of the AWMs and local PWVs were calculated and compared with ones without interpolation. The means of the NRMSEs for the AWMs and local PWVs based on the MTRI method with one interpolated sample decrease from 1.14% to 0.60% and 7.48% to 4.61%, respectively. Moreover, Bland-Altman analysis and coefficient of variation were used to validate the performance of the MTRI method based on the measured local PWVs of 30 healthy subjects. In conclusion, the reconstructive interpolation for the pulse wave estimation improves the accuracy and repeatability of the carotid local PWV measurement.

Double-Network Organohydrogels Toughened by Solvent Exchange.

Double-network hydrogels based on calcium alginate are extensively exploited. Unfortunately, their low strength and unstable constitution to open environments limit their application potential. Herein, a new type of double-network organohydrogel (OHG) is proposed. By solvent exchange, a stable physical network is established based on dimethyl sulfoxide (DMSO)-alginate in the presence of a polyacrylamide network. The DMSO content endows tunable mechanical properties, with a maximum tensile strength of ≈1.7 MPa. Importantly, the OHG shows much better environmental stability compared to the conventional double-network hydrogels. Due to the reversible association of hydrogen bonds, the OHG possesses some unique properties, including free-shapeability, shape-memory, and self-adhesion, that offers several promising ways to utilize alginate-based gels for wide applications.

Ultra-fast ultrasound blood flow velocimetry for carotid artery with deep learning.

Accurate measurement of blood flow velocity is important for the prevention and early diagnosis of atherosclerosis. However, due to the uncertainty of parameter settings, the autocorrelation velocimetry methods based on clutter filtering are prone to incorrectly filter out the near-wall blood flow signal, resulting in poor velocimetric accuracy. In addition, the Doppler coherent compounding acts as a low-pass filter, which also leads to low values of blood flow velocity estimated by the above methods. Motivated by this status quo, here we propose a deep learning estimator that combines clutter filtering and blood flow velocimetry based on the adaptive property of one-dimensional convolutional neural network (1DCNN). The estimator is operated by first extracting the blood flow signal from the original Doppler echo signal through an affine transformation of the 1D convolution, and then converting the extracted signal into the desired blood flow velocity using a linear transformation function. The effectiveness of the proposed method is verified by simulation as well as in vivo carotid artery data. Compared with typical velocimetry methods such as high-pass filtering (HPF) and singular value decomposition (SVD), the results show that the normalized root means square error (NRMSE) obtained by 1DCNN is reduced by 54.99 % and 53.50 % for forward blood flow velocimetry, and 70.99 % and 69.50 % for reverse blood flow velocimetry, respectively. Consistently, the in vivo measurements demonstrate that the goodness-of-fit of the proposed estimator is improved by 8.72 % and 4.74 % for five subjects. Moreover, the estimation time consumed by 1DCNN is greatly reduced, which costs only 2.91 % of the time of HPF and 12.83 % of the time of SVD. In conclusion, the proposed estimator is a better alternative to the current blood flow velocimetry, and is capable of providing more accurate diagnosis information for vascular diseases in clinical applications.

A Study on a Parameter Estimator for the Homodyned K Distribution Based on Table Search for Ultrasound Tissue Characterization.

OBJECTIVE: The homodyned K (HK) distribution is considered to be the most suitable distribution in the context of tissue characterization; therefore, the search for a rapid and reliable parameter estimator for HK distribution is important. METHODS: We propose a novel parameter estimator based on a table search (TS) for HK parameter estimates. The TS estimator can inherit the strength of conventional estimators by integrating various features and taking advantage of the TS method in a rapid and easy operation. Performance of the proposed TS estimator was evaluated and compared with that of XU (the estimation method based on X and U statistics) and artificial neural network (ANN) estimators. DISCUSSION: The simulation results revealed that the TS estimator is superior to the XU and ANN estimators in terms of normalized standard deviations and relative root mean squared errors of parameter estimation, and is faster. Clinical experiments found that the area under the receiver operating curve for breast lesion classification using the parameters estimated by the TS estimator could reach 0.871. CONCLUSION: The proposed TS estimator is more accurate, reliable and faster than the state-of-the-art XU and ANN estimators and has great potential for ultrasound tissue characterization based on the HK distribution.

Global trends and prospects in research of artificial cornea over past 20 years: a bibliometric and visualized analysis.

BACKGROUND: Many studies have been focused on the area of the artificial cornea. In our study, a bibliometric analysis was performed on the artificial cornea to identify the global key research fields and trends over the past 20 years. METHODS: Publications about artificial cornea were retrieved and downloaded from the Web of Science Core Collection from 2002 to 2021. CiteSpace and VOSviewer were used to analyze countries, institutions, authors, and related research areas. RESULTS: A total of 829 eligible publications were analyzed. The USA was the most productive country for the artificial cornea, followed by China and Canada. Harvard University was the most prolific institution in this field. Cornea published most of the studies in this area and Dohlman CH was the most cited author. CONCLUSIONS: Bibliometric analysis in our study first provides a general perspective on the artificial cornea, which can be helpful to further explore the issues in the rapidly developing area.

High-fidelity Cas13 variants for targeted RNA degradation with minimal collateral effects.

CRISPR-Cas13 systems have recently been used for targeted RNA degradation in various organisms. However, collateral degradation of bystander RNAs has limited their in vivo applications. Here, we design a dual-fluorescence reporter system for detecting collateral effects and screening Cas13 variants in mammalian cells. Among over 200 engineered variants, several Cas13 variants including Cas13d and Cas13X exhibit efficient on-target activity but markedly reduced collateral activity. Furthermore, transcriptome-wide off-targets and cell growth arrest induced by Cas13 are absent for these variants. High-fidelity Cas13 variants show similar RNA knockdown activity to wild-type Cas13 but no detectable collateral damage in transgenic mice or adeno-associated-virus-mediated somatic cell targeting. Thus, high-fidelity Cas13 variants with minimal collateral effects are now available for targeted degradation of RNAs in basic research and therapeutic applications.

Stability Analysis in Milling Based on the Localized Differential Quadrature Method.

Chatter stability analysis is an effective way to optimize the cutting parameters and achieve chatter-free machining. This paper proposes a milling chatter stability analysis method based on the localized differential quadrature method (LDQM), which has the advantages of simple principle, easy application, and high computational efficiency. The milling process, considering the regeneration effect, is modeled using linear periodic delay differential equations (DDE), then the state transition matrix during the adjacent cutting period is constructed based on the LDQM, and finally, the stability of the milling process is obtained based on the Floquet theory. The accuracy and computation efficiency of the proposed method are verified through two benchmark milling models. The simulation results demonstrate that the proposed method in this paper can accurately and quickly obtain the chatter stability lobe diagram (SLD) of the milling process, which will provide guidance for optimizing the process parameters.

[Genetic identification and sequence analysis of three individuals of rare ABO variant Bw subgroup].

OBJECTIVE: To identify and analysis three ABO variant Bw subtypes. METHODS: Serological assays were carried out to identify the ABO blood group of the proband. ABO gene was identified by Sanger sequencing. RESULTS: The genotype of three individuals are ABO*Bw.11/0.01.02, ABO*Bw.12/0.01.01, ABO*Bw.34/A1.02, receptively. Sequencing results showed that there were c.695T>C, c.278C>T, c.889G>A, resulting in variants in Leu232Pro, Pro93Leu and Glu297Lys, receptively. CONCLUSION: Bw11, Bw12 and Bw34 subgroups were identified, and gene testing can be used as a supplement to determine the ABO blood group subtypes.

Optimally filtering and matching processing for regional upstrokes to improve ultrasound transit time-based local PWV estimation.

BACKGROUND AND OBJECTIVE: Pulse wave velocity (PWV) is an important index for quantifying the elasticity of artery. Local PWV estimates based on ultrasonic transit time (TT) methods, however, are affected by the reflected waves and ultrasonic noise, biasing the spatiotemporal propagation of the time fiduciary point (TFP) positioning in the distension waveforms. In this study, an optimally filtering and matching processing for regional upstrokes is proposed to improve the ultrasound TT-based local PWV estimation. METHOD: (i) Smooth the pulse waves (PWs) using the Savitzky-Golay filter with one set of randomly combined parameters. (ii) An arbitrary region at the first beam upstroke of the smoothed PWs is selected as the curve template, and then matched with the upstrokes of other PWs by calculating the sum of square differences (SSD) between the template and matching regions to find its similar regions. (iii) Update the filter parameters and the template using the moth-flame optimization (MFO) feedback for computing the new SSD value. When the new SSD value is smaller than the historical one, the later will be replaced. (iv) Repeat the above steps until the MFO algorithm converges to the minimum SSD value. (v) Output the optimal filter parameters and the locations of regional curves corresponding to the minimum SSD value. Then the time delay of the PWs propagation can be detected by using the starting points of the regional curves as the TFPs. RESULTS: We conducted performance comparison with the advanced TT method through both simulation and clinical experiments. The results demonstrate that the proposed work observes considerable reductions on both the normalized root mean square error ± the standard deviation (from 6.73 ± 2.27% to 1.57 ± 0.72%) and the coefficient of variation (from 13.39% to 8.87%). CONCLUSIONS: The results of this study support that the proposed method may facilitate the early diagnosis and prevention of local arterial stiffness .

Association of Dietary Intake and Biomarker of α-Linolenic Acid With Incident Colorectal Cancer: A Dose-Response Meta-Analysis of Prospective Cohort Studies.

Background and Objective: There is keen interest in better understanding the impacts of alpha-linolenic acid (ALA), a plant-derived n-3 fatty acid, in ameliorating the development of cancer; however, results of several prospective cohorts present an inconsistent association between ALA intake and the incident colorectal cancer (CRC). We aimed to investigate the summary association of dietary intake and biomarkers of ALA with CRC risk based on the prospective cohorts. Methods: Pertinent prospective cohorts were identified in Cochrane Library, PubMed, and EMBASE from inception to February 2022. Study-specific risk ratios (RRs) with 95% confidence intervals (CIs) for comparing the top with the bottom quartiles of ALA levels were combined using a random-effects model. Nonlinear dose-response relationships of ALA levels in diet and blood with CRC risk were assessed using the restricted cubic spline models, respectively. Results: Over the duration of follow-up with a median of 9.3 years ranging from 1 to 28 years, 12,239 CRC cases occurred among 861,725 participants from 15 cohorts (11 studies on diet and 5 studies on biomarkers including 4 on blood and 1 on adipose tissue). The summary RR was 1.03 (95% CI: 0.97, 1.10; I2: 0.00%) for dietary intake and 0.83 (95% CI: 0.69, 0.99; I2: 0.00%) for biomarker. Each 0.1% increase in the levels of ALA in blood was associated with a 10% reduction in risk of CRC (summary RR: 0.90, 95% CI: 0.80, 0.99; I2: 38.60%), whereas no significant dose-response association was found between dietary intake of ALA and the incident CRC (p for non-linearity = 0.18; p for linearity = 0.24). Conclusions: Blood levels of ALA were inversely and linearly associated with the risk of CRC, which suggested that increased intake of ALA to improve circulating levels was beneficial for CRC prevention.

An Improved Parameter Estimator of the Homodyned K Distribution Based on the Maximum Likelihood Method for Ultrasound Tissue Characterization.

The homodyned K distribution (HK) can generally describe the ultrasound backscatter envelope statistics distribution with parameters that have specific physical meaning. However, creating robust and reliable HK parameter estimates remains a crucial concern. The maximum likelihood estimator (MLE) usually yields a small variance and bias in parameter estimation. Thus, two recent studies have attempted to use MLE for parameter estimation of HK distribution. However, some of the statements in these studies are not fully justified and they may hinder the application of parameter estimation of HK distribution based on MLE. In this study, we propose a new parameter estimator for the HK distribution based on the MLE (i.e., MLE1), which overcomes the disadvantages of conventional MLE of HK distribution. The MLE1 was compared with other estimators, such as XU estimator (an estimation method based on the first moment of the intensity and tow log-moments) and ANN estimator (an estimation method based on artificial neural networks). We showed that the estimations of parameters α and k are the best overall (in terms of the relative bias, normalized standard deviation, and relative root mean squared errors) using the proposed MLE1 compared with the others based on the simulated data when the sample size was N = 1000. Moreover, we assessed the usefulness of the proposed MLE1 when the number of scatterers per resolution cell was high (i.e., α up to 80) and when the sample size was small (i.e., N = 100), and we found a satisfactory result. Tests on simulated ultrasound images based on Field II were performed and the results confirmed that the proposed MLE1 is feasible and reliable for the parameter estimation from the ultrasonic envelope signal. Therefore, the proposed MLE1 can accurately estimate the HK parameters with lower uncertainty, which presents a potential practical value for further ultrasonic applications.

[Design and implementation for portable ultrasound-aided breast cancer screening system].

Early screening is an important means to reduce breast cancer mortality. In order to solve the problem of low breast cancer screening rates caused by limited medical resources in remote and impoverished areas, this paper designs a breast cancer screening system aided with portable ultrasound Clarius. The system automatically segments the tumor area of the B-ultrasound image on the mobile terminal and uses the ultrasound radio frequency data on the cloud server to automatically classify the benign and malignant tumors. Experimental results in this study show that the accuracy of breast tumor segmentation reaches 98%, and the accuracy of benign and malignant classification reaches 82%, and the system is accurate and reliable. The system is easy to set up and operate, which is convenient for patients in remote and poor areas to carry out early breast cancer screening. It is beneficial to objectively diagnose disease, and it is the first time for the domestic breast cancer auxiliary screening system on the mobile terminal.

Resolution of (R,S)-1-(4-methoxyphenyl)ethanol by lipase-catalyzed stereoselective transesterification and the process optimization.

An efficient lipase-catalyzed stereoselective transesterification reaction system was established for resolution of 1-(4-methoxyphenyl)ethanol (MOPE) enantiomers. A series of lipases were tested and compared. The immobilized lipase Novozym 40086 is selected as the best choice. The effects of organic solvent, acyl donor, time and temperature on substrate conversion (c), and optical purity of the remaining substrate (eeS ) were investigated. Response surface methodology and central composite design were employed to evaluate the effect of some important factors and to optimize the process. Under the optimized conditions including solvent of n-hexane, acyl donor of vinyl acetate, temperature of 35°C, substrate molar ratio of 1:6, enzyme dosage of 20 mg, and reaction time of 2.5 h, eeS of 99.87% with c of 56.71% is achieved. The use of alkane solvent and immobilized enzyme, the mild reaction conditions, and the reduced reaction time make the system promising in industrial application.

Ultrasound simulation of blood with different red blood cell aggregations and concentrations.

BACKGROUND: Considerable progress of ultrasound simulation on blood has enhanced the characterizing of red blood cell (RBC) aggregation. OBJECTIVE: A novel simulation method aims at modeling the blood with different RBC aggregations and concentrations is proposed. METHODS: The modeling process is as follows: (i) A three-dimensional scatterer model is first built by a mapping with a Hilbert space-filling curve from the one-dimensional scatterer distribution. (ii) To illustrate the relationship between the model parameters and the RBC aggregation level, a variety of blood samples are prepared and scanned to acquire their radiofrequency signals in-vitro. (iii) The model parameters are determined by matching the Nakagami-distribution characteristics of envelope signals simulated from the model with those measured from the blood samples. RESULTS: Nakagami metrics m estimated from 15 kinds of blood samples (hematocrits of 20%, 40%, 60% and plasma concentrations of 15%, 30%, 45%, 60%, 75%) are compared with metrics estimated by their corresponding models (each with different eligible parameters). Results show that for the three hematocrit levels, the mean and standard deviation of the root-mean-squared deviations of m are 0.27 ± 0.0026, 0.16 ± 0.0021, 0.12 ± 0.0018 respectively. CONCLUSION: The proposed simulation model provides a viable data source to evaluate the performance of the ultrasound-based methods for quantifying RBC aggregation.

Programmable RNA editing with compact CRISPR-Cas13 systems from uncultivated microbes.

Competitive coevolution between microbes and viruses has led to the diversification of CRISPR-Cas defense systems against infectious agents. By analyzing metagenomic terabase datasets, we identified two compact families (775 to 803 amino acids (aa)) of CRISPR-Cas ribonucleases from hypersaline samples, named Cas13X and Cas13Y. We engineered Cas13X.1 (775 aa) for RNA interference experiments in mammalian cell lines. We found Cas13X.1 could tolerate single-nucleotide mismatches in RNA recognition, facilitating prophylactic RNA virus inhibition. Moreover, a minimal RNA base editor, composed of engineered deaminase (385 aa) and truncated Cas13X.1 (445 aa), exhibited robust editing efficiency and high specificity to induce RNA base conversions. Our results suggest that there exist untapped bacterial defense systems in natural microbes that can function efficiently in mammalian cells, and thus potentially are useful for RNA-editing-based research.