Efficient and Stable Proton Exchange Membrane Water Electrolysis Enabled by Stress Optimization.

Proton exchange membrane water electrolysis (PEMWE) is a promising solution for the conversion and storage of fluctuating renewable energy sources. Although tremendously efficient materials have been developed, commercial PEMWE products still cannot fulfill industrial demands regarding efficiency and stability. In this work, we demonstrate that the stress distribution, a purely mechanical parameter in electrolyzer assembly, plays a critical role in overall efficiency and stability. The conventional cell structure, which usually adopts a serpentine flow channel (S-FC) to deliver and distribute reactants and products, resulted in highly uneven stress distribution. Consequently, the anode catalyst layer (ACL) under the high stress region was severely deformed, whereas the low stress region was not as active due to poor electrical contact. To address these issues, we proposed a Ti mesh flow channel (TM-FC) with gradient pores to reduce the stress inhomogeneity. Consequently, the ACL with TM-FC exhibited 27 mV lower voltage initially and an 8-fold reduction in voltage degradation rate compared to that with S-FC at 2.0 A/cm2. Additionally, the applicability of the TM-FC was demonstrated in cross-scale electrolyzers up to 100 kW, showing a voltage increase of only 20 mV (accounting for less than 2% of overall voltage) after three orders of magnitude scaleup.

Identification and characterization of novel genetic variants in the first Chinese family of mucopolysaccharidosis IIIC (Sanfilippo C syndrome).

Mucopolysaccharidosis type IIIC (MPS IIIC) is one of inherited lysosomal storage disorders, caused by deficiencies in lysosomal hydrolases degrading acidic mucopolysaccharides. The gene responsible for MPS IIIC is HGSNAT, which encodes an enzyme that catalyses the acetylation of the terminal glucosamine residues of heparan sulfate. So far, few studies have focused on the genetic landscape of MPS IIIC in China, where IIIA and IIIB were the major subtypes. In this study, we utilized whole-exome sequencing (WES) to identify novel compound heterozygous variants in the HGSNAT gene from a Chinese patient with typical MPS IIIC symptoms: c.743G>A; p.Gly248Glu and c.1030C>T; p.Arg344Cys. We performed in silico analysis and experimental validation, which confirmed the deleterious pathogenic nature of both variants, as evidenced by the loss of HGSNAT activity and failure of lysosomal localization. To the best of our knowledge, the MPS IIIC is first confirmed by clinical, biochemical and molecular genetic findings in China. Our study thus expands the spectrum of MPS IIIC pathogenic variants, which is of importance to dissect the pathogenesis and to carry out clinical diagnosis of MPS IIIC. Moreover, this study helps to depict the natural history of Chinese MPS IIIC populations.

Bioinspired Jumping Soft Actuators of the Liquid Crystal Elastomer Enabled by Photo-Mechanical Coupling.

Jumping, a fundamental survival behavior observed in organisms, serves as a vital mechanism for adapting to the surrounding environment and overcoming significant obstacles within a given terrain. Here, we present a light-controlled soft jumping actuator inspired by asphondylia, which employs a closed-loop structure and utilizes a liquid crystal elastomer (LCE). Photo-mechanical coupling highlights the significant influence of the light source on the actuator's jumping behavior. Manipulating the light intensity, the relative position of stimulus and light lock, and the concentration of disperse red 1 (DR1) allows precise control over both the maximum take-off velocity and jump height. Furthermore, tailoring the size of the LCE actuator offers a means of regulating jumping behavior. Upon exposure to 460 nm LED irradiation, our actuator achieves remarkable performance, with a maximum jumping height of 10 body length (BL) and take-off velocity of 62 BL/s. These actuators accumulate and rapidly release energy, enabling the effective transportation of microcargos across substantial distances. Our research yields valuable insights into the realm of soft robotics, underscoring the pivotal importance of photo-mechanical coupling in the field of soft robotics, thereby serving as a catalyst for inspiring continued exploration of agile and capable systems by prestoring elastic energy.

Characterization of the circRNA Landscape in Interleukin-4 Induced Anti-Inflammatory Microglia.

Microglia are resident innate immune cells that play an essential role in the development and surveillance of the central nervous system as well as the shared pathogenesis of neurodegenerative diseases. Microglia rapidly respond to multiple inflammatory stimuli and activate towards different phenotypes, such as pro-inflammatory and anti-inflammatory phenotypes. Cytokines, epigenetic and long non-coding RNA modulations have been shown to regulate microglial activation; however, the role of circRNAs in microglia-mediated neuroinflammation remains elusive. Here, we performed circRNA sequencing in IL-4-treated anti-inflammatory microglia and discovered 120 differentially expressed circRNAs. We systemically verified the identities of circRNAs by assays of PCR, RNase R treatment and fluorescent in situ hybridization (FISH), among others. We found that circAdgre1 promoted IL-4-induced anti-inflammatory responses and further conferred neuroprotective effects upon lipopolysaccharide (LPS) stimuli. Taken together, our results show that circRNAs might be possible therapeutic targets for microglia-mediated neuroinflammation and neurodegenerative diseases.

Citrulline inhibits LPS-induced pyroptosis of RAW264.7 macrophages through NF-κB signaling pathway.

BACKGROUND: The aim of this study was to investigate the effect of citrulline on the pyroptosis of mouse macrophage RAW264.7 and the mechanism. We investigated the effect of citrulline on pyroptosis of RAW264.7 cell induced by lipopolysaccharide (LPS), and the modulation of nuclear factor-kappaB (NF-κB) signaling. METHODS: Pyroptosis was evaluated using flow cytometry and caspase-1/sytox double staining. Cell counting kit-8 assay was performed to evaluate cell viability. RESULTS: Citrulline promoted cell viability and inhibited the pyroptosis of RAW264.7 cell stimulated by LPS. Furthermore, citrulline inactivated NF-κb/p65 signaling pathway by suppressing p65 nuclear translocation induced by LPS. An NF-κb signaling pathway activator, betulinic acid, reversed the inhibition of pyroptosis induced by citrulline. CONCLUSION: Citrulline inhibited LPS-induced pyrophosis, which may be closely related to the inactivation of NF-κB/p65 signaling pathway.

Effects of ammonia on morphological characteristics and nanostructure of soot in the combustion of diesel surrogate fuels.

The morphological characteristics and nanostructure of soot particles in pure n-heptane (C7H16) and n-heptane/ammonia co-flow diffusion flames were analyzed and compared using thermophoretic sampling and transmission electron microscopy (TEM) observation combining with quantitative image information extraction methods. The results showed that the overall formation and evolution of soot particles in NH3-doped n-heptane flames along the flame centerline were similar with that without NH3-doping. However, compared to n-heptane flame, the peak average diameter of primary soot particles and the peak gyration radius of soot aggregates in NH3-doped flames were reduced by about 45% and 37%, respectively, which indicated that the growth of both primary soot particles via surface reaction/condensation and soot aggregates via coagulation were significantly decreased. Meanwhile, the fractal dimension of soot aggregates was lower with NH3 addition as the structure of soot aggregates was looser and tended to be more chain-like. After NH3 doping, the peak average fringe length inside soot particles was decreased by 13%, and the inter-fringe spacing and tortuosity of soot were increased by 8% and 3%, respectively. This represented a more disordered microcrystal structure and lower degree of graphitization of soot particles, meaningfully indicating a higher oxidation reactivity.

The heterogeneity of microglial activation and its epigenetic and non-coding RNA regulations in the immunopathogenesis of neurodegenerative diseases.

Microglia are resident immune cells in the brain and play a central role in the development and surveillance of the nervous system. Extensive gliosis is a common pathological feature of several neurodegenerative diseases, such as Alzheimer's disease (AD), the most common cause of dementia. Microglia can respond to multiple inflammatory insults and later transform into different phenotypes, such as pro- and anti-inflammatory phenotypes, thereby exerting different functions. In recent years, an increasing number of studies based on both traditional bulk sequencing and novel single-cell/nuclear sequencing and multi-omics analysis, have shown that microglial phenotypes are highly heterogeneous and dynamic, depending on the severity and stage of the disease as well as the particular inflammatory milieu. Thus, redirecting microglial activation to beneficial and neuroprotective phenotypes promises to halt the progression of neurodegenerative diseases. To this end, an increasing number of studies have focused on unraveling heterogeneous microglial phenotypes and their underlying molecular mechanisms, including those due to epigenetic and non-coding RNA modulations. In this review, we summarize the epigenetic mechanisms in the form of DNA and histone modifications, as well as the general non-coding RNA regulations that modulate microglial activation during immunopathogenesis of neurodegenerative diseases and discuss promising research approaches in the microglial era.

The Double-Faceted Role of Leucine-Rich Repeat Kinase 2 in the Immunopathogenesis of Parkinson's Disease.

Leucine-rich repeat kinase 2 (LRRK2) is one of the most common causative genes in Parkinson's disease (PD). The complex structure of this multiple domains' protein determines its versatile functions in multiple physiological processes, including migration, autophagy, phagocytosis, and mitochondrial function, among others. Mounting studies have also demonstrated the role of LRRK2 in mediating neuroinflammation, the prominent hallmark of PD, and intricate functions in immune cells, such as microglia, macrophages, and astrocytes. Of those, microglia were extensively studied in PD, which serves as the resident immune cell of the central nervous system that is rapidly activated upon neuronal injury and pathogenic insult. Moreover, the activation and function of immune cells can be achieved by modulating their intracellular metabolic profiles, in which LRRK2 plays an emerging role. Here, we provide an updated review focusing on the double-faceted role of LRRK2 in regulating various cellular physiology and immune functions especially in microglia. Moreover, we will summarize the latest discovery of the three-dimensional structure of LRRK2, as well as the function and dysfunction of LRRK2 in immune cell-related pathways.

Identification and characterization of two novel noncoding tyrosinase (TYR) gene variants leading to oculocutaneous albinism type 1.

Oculocutaneous albinism type 1 (OCA1), resulting from pathogenic variants in the tyrosinase (TYR) gene, refers to a group of phenotypically heterogeneous autosomal recessive disorders characterized by a partial or a complete absence of pigment in the skin/hair and is also associated with common developmental eye defects. In this study, we identified two novel compound heterozygous TYR variants from a Chinese hypopigmentary patient by whole-exome sequencing. Specifically, the two variants were c.-89T>G, located at the core of the initiator E-box (Inr E-box) of the TYR promoter, and p.S16Y (c.47C>A), located within the signal sequence. We performed both in silico analysis and experimental validation and verified these mutations as OCA1 variants that caused either impaired or complete loss of function of TYR. Mechanistically, the Inr E-box variant dampened TYR binding to microphthalmia-associated transcription factor, a master transcriptional regulator of the melanocyte development, whereas the S16Y variant contributed to endoplasmic reticulum retention, a common and principal cause of impaired TYR activity. Interestingly, we found that the Inr E-box variant creates novel protospacer adjacent motif sites, recognized by nucleases SpCas9 and SaCas9-KKH, respectively, without compromising the functional TYR coding sequence. We further used allele-specific genomic editing by CRISPR activation to specifically target the variant promoter and successfully activated its downstream gene expression, which could lead to potential therapeutic benefits. In conclusion, this study expands the spectrum of TYR variants, especially those within the promoter and noncoding regions, which can facilitate genetic counseling and clinical diagnosis of OCA1.

Study on the removal characteristics of different air pollution control devices for condensable particulate matter in coal-fired power plants.

This study reports the emissions of condensable particulate matter (CPM) and filterable particulate matter (FPM) in two coal-fired power plants with different air pollution control devices (APCDs). The mechanisms of CPM removed by existing APCDs in coal-fired power plants were explored, and a series of analyses were also carried out on the composition and characteristics of CPM. The results show that the removal efficiencies to CPM by electrostatic-bag-precipitator (EBP) and ESP are 77.34% and 79.23%, respectively, so the difference is not obvious because the interception filtration mechanisms of baghouses for CPM have less effect on CPM compared to FPM. The mechanism of EBP/ESP to remove CPM is mainly electrostatic adsorption and FPM's adsorption. The concentration of CPM decreases when passing through WFGD. However, the WESP can increase the CPM in different ways. For example, the pollution of the circulation of the flushing fluid may cause the increase of CPM. In addition, CPM mainly includes three parts. The first part is organic fractions such as alkanes and esters; the second is the water-soluble ions that include SO42-, NH4+, and Cl-; and the third is Na, Ca, and other minerals. The research in this study is helpful to understand the impact of existing APCDs in coal-fired power plants on CPM and the sources of CPM.

A Step-by-Step Refined Strategy for Highly Efficient Generation of Neural Progenitors and Motor Neurons from Human Pluripotent Stem Cells.

Limited access to human neurons, especially motor neurons (MNs), was a major challenge for studying neurobiology and neurological diseases. Human pluripotent stem cells (hPSCs) could be induced as neural progenitor cells (NPCs) and further multiple neural subtypes, which provide excellent cellular sources for studying neural development, cell therapy, disease modeling and drug screening. It is thus important to establish robust and highly efficient methods of neural differentiation. Enormous efforts have been dedicated to dissecting key signalings during neural commitment and accordingly establishing reliable differentiation protocols. In this study, we refined a step-by-step strategy for rapid differentiation of hPSCs towards NPCs within merely 18 days, combining the adherent and neurosphere-floating methods, as well as highly efficient generation (~90%) of MNs from NPCs by introducing refined sets of transcription factors for around 21 days. This strategy made use of, and compared, retinoic acid (RA) induction and dual-SMAD pathway inhibition, respectively, for neural induction. Both methods could give rise to highly efficient and complete generation of preservable NPCs, but with different regional identities. Given that the generated NPCs can be differentiated into the majority of excitatory and inhibitory neurons, but hardly MNs, we thus further differentiate NPCs towards MNs by overexpressing refined sets of transcription factors, especially by adding human SOX11, whilst improving a series of differentiation conditions to yield mature MNs for good modeling of motor neuron diseases. We thus refined a detailed step-by-step strategy for inducing hPSCs towards long-term preservable NPCs, and further specified MNs based on the NPC platform.

Using artificial neural network in predicting the key fruit quality of loquat.

The formation and regulation of loquat fruit quality have always been an important research field to improve fruit quality, commodities, and market value. Fruit size, soluble solids content, and titratable acid content represent the most important quality factors in loquat. Mineral nutrients in abundance or deficiency are among the most important key factor that affect fruit quality. In the present study, we use artificial neural network (ANN) to explore the effects of mineral nutrients in soil and leaves on the key fruit quality of loquat. The results show that the ANN model with the structure of 12-12-1 can predict the single fruit weight with the highest accuracy (R 2 = .91), the ANN model with the structure of 10-11-1 can predict the soluble solid content with the highest accuracy (R 2 = .91), and the ANN model with the structure of 9-10-1 can predict the titratable acid content with the highest accuracy (R 2 = .95). Meanwhile, we also conduct sensitivity analysis to analyze the relative contribution of mineral nutrients in soils and leaves to determine of the key fruit quality. In terms of relative contribution, Ca, Fe, and Mg content in soils and Zn, K, and Ca content in leaves contribute relatively largely to a single fruit weight, Mn, Fe, and Mg content in soils and the N content in leaves contribute relatively largely to the soluble solid content, and the P, Ca, N, Mg, and Fe in leaves contribute relatively largely to the titratable acid content of loquat. The established artificial neural network prediction models can improve the quality of loquat fruit by optimizing the content of mineral elements in soils and leaves.

Rasagiline combined with levodopa therapy versus levodopa monotherapy for patients with Parkinson's disease: a systematic review.

OBJECTIVE: The aim of this report was to systematically evaluate the efficacy and safety of rasagiline (R) plus levodopa (L) (R + L) for the treatment of Parkinson's disease (PD) compared with that of L monotherapy, in order to provide a reference resource for rational drug use. METHODS: Randomized controlled trials (RCTs) of R + L for PD published up to September 2018 were searched. Sensitivity analyses were also performed. RESULTS: Fourteen RCTs with 2531 participants were included. Compared with L monotherapy, the pooled effects of R + L combination therapy on unified Parkinson's disease rating scale (UPDRS) score were (SMD - 0.50, 95% CI - 0.70 to - 0.30, P < 0.00001) for UPDRS motor score, (SMD - 0.59, 95% CI - 0.79 to - 0.39, P < 0.00001) for UPDRS activities of daily living (ADL) score, (SMD - 0.65, 95% CI - 0.81 to - 0.49, P < 0.00001) for UPDRS total score. R + L combination therapy was better than L monotherapy in reducing daily off-time (SMD - 1.15, 95% CI - 2.13 to - 0.17, P = 0.02), but there was a statistically nonsignificant result in daily on-time increase (SMD 1.39, 95% CI - 0.69 to 3.48, P = 0.19). There were no statistical differences in number of adverse events (OR 1.33, 95% CI 0.97 to 1.82, P = 0.07) and number of dropout (OR 0.88, 95% CI 0.65 to 1.19, P = 0.39) between R + L combination therapy and L monotherapy. CONCLUSIONS: R + L combination therapy was superior to L monotherapy for improvement of UPDRS scores and off-time in PD patients. Moreover, R + L combination therapy and L monotherapy were similar in terms of safety and tolerability.

Transcriptome analysis of Chinese bayberry (Myrica rubra Sieb. et Zucc.) fruit treated with heat and 1-MCP.

Chinese bayberry (Myrica rubra Sieb. et Zucc.) is a typical fruit tree grown in the hilly region of Southern China. The fruit is sensitive to storage and transportation conditions and presents a major problem in its commercialization. The present study was conducted to investigate the regulation of gene expression involved in plant hormone signaling pathway in the Chinese bayberry with different treatments of heat and 1-methylcyclopene (1-MCP) during postharvest storage. In one treatment group (HM group), we exposed Chinese bayberry fruit to 48 °C for 10 min and then sealed them in a desiccator with 5 µl·L-1 of 1-MCP for 24 h at 20 °C, followed by storage at 10 °C. Another group (CK group) was directly stored at 10 °C without any prior treatment. Samples of fruit were collected every three days, at 3, 6, 9, 12 and 15 d (CK3, CK6, CK9, CK12 and CK15; and HM3, HM6, HM9, HM12, and HM15, respectively). The decay index of fruits in the CK group increased after six days of storage but did not increase until nine days of storage in the HM group. Superoxide dismutase (SOD) activity in the CK group was shown a downtrend during storage, and almost no fluctuation from six days. In the HM group, SOD activity increased after three days, but decreased sharply after six days storage. Besides, peroxidase (POD) and catalase (CAT) activities were shown the similar trend during the storage, both of them first increased and then decreased form the six days of storage. These physiological data indicated that the sixth day is a crucial time during the storage of Chinese bayberry treated with heat and 1-MCP. Therefore, the transcriptome libraries were constructed from CK0, CK6, HM6 group, respectively. The analysis of top 20 KEGG pathways showed that most differentially expressed genes were involved in the biosynthesis of secondary metabolites, particularly flavonoids and flavanols biosynthesis, in CK0 vs. CK6 and CK0 vs. HM6. However, the top three KEGG pathways in CK6 vs. HM6 were the ribosome, RNA transport and endocytosis during the storage. Expression of six ethylene receptor (ETR) genes and four ethylene-responsive transcription factor (ERF) genes were activated at transcriptional level during the postharvest stage and were decreased by heat and 1-MCP treatment, and serine/threonine-protein kinase 1 (CTR1) was also repressed by treatment. Abscisic acid (ABA) -responsive element binding factor (ABF) gene, auxin-responsive GH3 gene and transcription factor MYC2 gene also showed similar expression pattern with ethylene pathway genes. These results might improve our understanding of the mechanisms of heat and 1-MCP inhibition of fruit postharvest physiology and prolongation of fruit shelf life.

Integrating proteomic and transcriptomic analyses of loquat (Eriobotrya japonica Lindl.) in response to cold stress.

The expression levels of many genes and the related proteins change and regulate physiological and metabolic processes that help the plant survive harsh environmental conditions under cold stress. Damage due to cold and freezing conditions often causes dynamic loss of loquat fruits in cultivated parts of northern China. To illustrate the mechanism of cold tolerance in the loquat, we combined the transcriptomic analysis with isobaric tags for relative and absolute quantification (iTRAQ) and RNA sequencing (RNA-Seq) data from loquat leaves under 4 °C treatment. The results showed 122,081 genes and 1210 differentially expressed genes (DEGs), while only 4582 proteins and 300 differential proteins (DEPs) were identified. Functional annotation and Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway analysis indicated that metabolic pathways and biosynthesis of secondary metabolites were the two most common pathways in transcriptional and translational processes in this study. Comparison analysis of the transcriptomic and proteomic profiles, only 27 of 3620 genes were found to be shared both in DEGs and DEPs. Further validation with Real-Time Quantitative RT-PCR analysis showed that the genes expression of NADP-dependent D-sorbitol-6-phosphate dehydrogenase, anthocyanin synthase and phenylalanine ammonia-lyase were consistent with the pattern of transcriptome profile, which suggested that these three genes might play vital roles in cold tolerance in loquat.

A new strategy for estimating two-locus recombination fractions under some natural inequality restrictions.

Linkage analysis is now being widely used to map markers on each chromosome in the human genome, to map genetic diseases, and to identify genetic forms of common diseases. Two-locus linkage analysis and multi-locus analysis have been investigated comprehensively, and many computer programs have been developed to perform linkage analysis. Yet there exists a shortcoming in traditional methods, i.e., the parameter space of two-locus recombination fractions has not been emphasized sufficiently in the usual analyses. In this paper, we propose a new strategy for estimating the two-locus recombination fractions based on data of backcross family in the framework of some natural and necessary parameter restrictions. The new strategy is based on a restricted projection algorithm, which can provide fast reasonable estimates of recombination fraction, and can therefore serve as a superior alternative algorithm. Results obtained from both real and simulated data indicate that the new algorithm performs well in the estimation of recombination fractions and outperforms current methods.

A robust approach based on Weibull distribution for clustering gene expression data.

BACKGROUND: Clustering is a widely used technique for analysis of gene expression data. Most clustering methods group genes based on the distances, while few methods group genes according to the similarities of the distributions of the gene expression levels. Furthermore, as the biological annotation resources accumulated, an increasing number of genes have been annotated into functional categories. As a result, evaluating the performance of clustering methods in terms of the functional consistency of the resulting clusters is of great interest. RESULTS: In this paper, we proposed the WDCM (Weibull Distribution-based Clustering Method), a robust approach for clustering gene expression data, in which the gene expressions of individual genes are considered as the random variables following unique Weibull distributions. Our WDCM is based on the concept that the genes with similar expression profiles have similar distribution parameters, and thus the genes are clustered via the Weibull distribution parameters. We used the WDCM to cluster three cancer gene expression data sets from the lung cancer, B-cell follicular lymphoma and bladder carcinoma and obtained well-clustered results. We compared the performance of WDCM with k-means and Self Organizing Map (SOM) using functional annotation information given by the Gene Ontology (GO). The results showed that the functional annotation ratios of WDCM are higher than those of the other methods. We also utilized the external measure Adjusted Rand Index to validate the performance of the WDCM. The comparative results demonstrate that the WDCM provides the better clustering performance compared to k-means and SOM algorithms. The merit of the proposed WDCM is that it can be applied to cluster incomplete gene expression data without imputing the missing values. Moreover, the robustness of WDCM is also evaluated on the incomplete data sets. CONCLUSIONS: The results demonstrate that our WDCM produces clusters with more consistent functional annotations than the other methods. The WDCM is also verified to be robust and is capable of clustering gene expression data containing a small quantity of missing values.

MicroRNA regulation constrains the organization of target genes on mammalian chromosomes.

The regulation of microRNAs (miRNAs) is a complicated process requiring a large number of molecular events to be coordinated in both space and time. It is not known whether this complicated regulation process constrains the organization of target genes on mammalian chromosomes. We performed a genome-wide analysis to provide a better picture of chromosomal organization of miRNA target genes. Our results showed clustering of the target genes (TGs) of miRNAs on mammalian chromosomes, and further revealed that the particular gene organization is constrained by miRNA regulation. The clustering pattern of TGs provides an insight into the complexity of miRNA regulation.

Expression of the human hepatitis B virus large surface antigen gene in transgenic tomato plants.

The original hepatitis B virus (HBV) large surface antigen gene was synthesized. In order to optimize the expression of this gene in tomato plants, the tobacco pathogenesis-related protein S signal peptide was fused to the 5' end of the modified gene and the sequence encoding amino acids S, E, K, D, E, and L was placed at the 3' end. The gene encoding the modified HBV large surface antigen under the control of a fruit-specific promoter was constructed and expressed in transgenic tomato plants. The expression of the antigen from transgenic plants was confirmed by PCR and reverse transcriptase PCR. Enzyme-linked immunoassays using a monoclonal antibody directed against human serum-derived HBsAg revealed that the maximal level of HBsAg was about 0.02% of the soluble protein in transgenic tomato fruit. The amount of HBsAg in mature fruits was found to be 65- to 171-fold larger than in small or medium fruits and leaf tissues. Examination of transgenic plant samples by transmission electron microscopy proved that HBsAg had been expressed and had accumulated. The HBsAg protein was capable of assembling into capsomers and virus-like particles. To our knowledge, this is the first time the HBV large surface antigen has been expressed in plants. This work suggests the possibility of producing a new alternative vaccine for human HBV.