Sperm-origin paternal effects on root stem cell niche differentiation.

Fertilization introduces parental genetic information into the zygote to guide embryogenesis. Parental contributions to postfertilization development have been discussed for decades, and the data available show that both parents contribute to the zygotic transcriptome, suggesting a paternal role in early embryogenesis1-6. However, because the specific paternal effects on postfertilization development and the molecular pathways underpinning these effects remain poorly understood, paternal contribution to early embryogenesis and plant development has not yet been adequately demonstrated7. Here our research shows that TREE1 and its homologue DAZ3 are expressed exclusively in Arabidopsis sperm. Despite presenting no evident defects in sperm development and fertilization, tree1 daz3 unexpectedly led to aberrant differentiation of the embryo root stem cell niche. This defect persisted in seedlings and disrupted root tip regeneration, comparable to congenital defects in animals. TREE1 and DAZ3 function by suppression of maternal RKD2 transcription, thus mitigating the detrimental maternal effects from RKD2 on root stem cell niche. Therefore, our findings illuminate how genetic deficiencies in sperm can exert enduring paternal effects on specific plant organ differentiation and how parental-of-origin genes interact to ensure normal embryogenesis. This work also provides a new concept of how gamete quality or genetic deficiency can affect specific plant organ formation.

Impacts of pilot carbon emission trading policies on urban environmental pollution: Evidence from China.

The trading of carbon emissions is a crucial regulatory method to address environmental pollution issues. This study takes China's carbon emission trading pilot policy established in 2013 as a quasi-natural experiment and uses the DID model to empirically test the urban panel data from 2006 to 2019. The results show that the carbon emission trading pilot policy can effectively reduce urban environmental pollution, and this effect is more noticeable in mid-western cities, northern cities, cities with fewer resources, and large-scale cities. In addition, to address the urban environmental pollution problem through this policy, the government is encouraged to raise its environmental protection awareness and put more effort into the innovation of technology. In general, this study uses carbon emission trading policies from China to confirm that market-based incentive environmental regulation tools can effectively reduce environmental pollution in urban areas. These findings can provide more theoretical support and empirical evidence for the government to use mechanisms of the market to effectively solve pollution problems, improve ecological environment quality, and accelerate the realization of green economy.

Enhancing polyethylene degradation: a novel bioprocess approach using Acinetobacter nosocomialis pseudo-resting cells.

Despite the discovery of several bacteria capable of interacting with polymers, the activity of the natural bacterial isolates is limited. Furthermore, there is a lack of knowledge regarding the development of bioprocesses for polyethylene (PE) degradation. Here, we report a bioprocess using pseudo-resting cells for efficient degradation of PE. The bacterial strain Acinetobacter nosocomialis was isolated from PE-containing landfills and characterized using low-density PE (LDPE) surface oxidation when incubated with LDPE. We optimized culture conditions to generate catalytic pseudo-resting cells of A. nosocomialis that are capable of degrading LDPE films in a bioreactor. After 28 days of bioreactor operation using pseudo-resting cells of A. nosocomialis, we observed the formation of holes on the PE film (39 holes per 217 cm2, a maximum diameter of 1440 µm). This study highlights the potential of bacteria as biocatalysts for the development of PE degradation processes. KEY POINTS: • New bioprocess has been proposed to degrade polyethylene (PE). • Process with pseudo-resting cells results in the formation of holes in PE film. • We demonstrated PE degradation using A. nosocomialis as a biocatalyst.

Efforts to install a heterologous Wood-Ljungdahl pathway in Clostridium acetobutylicum enable the identification of the native tetrahydrofolate (THF) cycle and result in early induction of solvents.

Here, we report the construction of a Clostridium acetobutylicum strain ATCC 824 (pCD07239) by heterologous expression of carbonyl branch genes (CD630_0723∼CD630_0729) from Clostridium difficile, aimed at installing a heterologous Wood-Ljungdahl pathway (WLP). As part of this effort, in order to validate the methyl branch of the WLP in the C. acetobutylicum, we performed 13C-tracing analysis on knockdown mutants of four genes responsible for the formation of 5-methyl-tetrahydrofolate (5-methyl-THF) from formate: CA_C3201, CA_C2310, CA_C2083, and CA_C0291. While C. acetobutylicum 824 (pCD07239) could not grow autotrophically, in heterotrophic fermentation, it began producing butanol at the early growth phase (OD600 of 0.80; 0.162 g/L butanol). In contrast, solvent production in the parent strain did not begin until the early stationary phase (OD600 of 7.40). This study offers valuable insights for future research on biobutanol production during the early growth phase.

Complete Genome Sequence of Acinetobacter nosocomialis GNU001, Isolated from a Plastic-Containing Landfill.

Due to the hazard of plastic waste exposed to the environment, microorganisms capable of degrading different polymeric pollutants have gained attention. Here, we report the complete genome sequence of Acinetobacter nosocomialis GNU001, which was isolated from a landfill. The genome was composed of a circular chromosome of 3,850,149 bp and a plasmid.

Environmental toxicity and decomposition of polyethylene.

In the more than 100 years since the invention of plastics, various plastic polymers have been developed that exhibit different characteristics and have been widely used in production and life. In 2020 alone, nearly 400 million tons of plastics were produced globally. However, while plastic such as polyethylene brings us convenience, it also threatens environmental sustainability and human health. Due to insufficient recycling efficiency, millions of tons of polyethylene pollutants accumulate in terrestrial or marine environments each year. Polyethylene is elastic, chemically stable, and non-biodegradable, and the traditional disposal methods include landfilling and incineration. These methods are costly, unsustainable, and further increase the burden on the environment. Therefore, recent research has increasingly focused on the biodegradation of polyethylene. In this work, we briefly summarized polyethylene's properties and environmental toxicity. We also reviewed the recent advances in the biodegradation of polyethylene with a summary of traditional abiotic methods. Finally, we proposed a brief research direction in polyethylene study with the aspect of environmental toxicology and industrial applications of decomposition technology.

DNA Shuffling of aprE Genes to Increase Fibrinolytic Activity and Thermostability.

Four aprE genes encoding alkaline serine proteases from B. subtilis strains were used as template genes for family gene shuffling. Shuffled genes obtained by DNase I digestion followed by consecutive primerless and regular PCR reactions were ligated with pHY300PLK, an E. coli-Bacillus shuttle vector. The ligation mixture was introduced into B. subtilis WB600 and one transformant (FSM4) showed higher fibrinolytic activity. DNA sequencing confirmed that the shuffled gene (aprEFSM4) consisted of DNA mostly originated from either aprEJS2 or aprE176 in addition to some DNA from either aprE3-5 or aprESJ4. Mature AprEFSM4 (275 amino acids) was different from mature AprEJS2 in 4 amino acids and mature AprE176 in 2 amino acids. aprEFSM4 was overexpressed in E. coli BL21 (DE3) by using pET26b(+) and recombinant AprEFSM4 was purified. The optimal temperature and pH of AprEFSM4 were similar to those of parental enzymes. However, AprEFM4 showed better thermostability and fibrinogen hydrolytic activity than the parental enzymes. The results indicated that DNA shuffling could be used to improve fibrinolytic enzymes from Bacillus sp. for industrial applications.

High cortisol and the risk of Alzheimer disease: A protocol for systematic review and meta-analysis.

BACKGROUND: Morning cortisol levels have been reported to be elevated among patients with Alzheimer disease (AD). We perform a protocol for systematic review and meta-analysis to assess morning central or peripheral cortisol levels in AD patients as compared with cognitively normal individuals. METHODS: Studies were identified through systematic searches in August 2021 with no restrictions on date and time, language, and publication status using the following bibliographic databases: Embase, Medline, PubMed, Web of Science, Science Direct, and the Cochrane Library. Studies were identified using search terms related to cortisol, Alzheimer disease, and cognitive impairment. The study quality of included papers was evaluated using the "National Institutes of Health (NIH) quality assessment tool for observational cohort and cross-sectional studies." Statistical analyses were performed using Stata (version 14, StataCorp, College Station, TX). RESULTS: The findings of this study will be submitted to peer-reviewed journals for publication. CONCLUSION: Morning cortisol was elevated in AD patients and may have diagnostic and prognostic values for AD.

H2O2 response gene 1/2 are novel sensors or responders of H2O2 and involve in maintaining embryonic root meristem activity in Arabidopsis thaliana.

Signal molecule hydrogen peroxide (H2O2) plays critical roles in various processes of plant development. However, H2O2 signaling network, especially the responders that sense and respond to the H2O2 signal remain largely unknown. Here we report two homologous genes H2O2 Response Gene 1 and 2 (HRG1/2) in Arabidopsis that could quickly respond to exogenous or endogenous H2O2. Knockdown of HRG1/2 facilitated seed germination while overexpression of HRG1/2 greatly retarded seed germination. ROS level in HRG1 overexpression roots was significantly lower than that in HRG1/2 mutants after H2O2 treatment. The expression level of enzymatic antioxidant DHAR3 was upregulated in HRG1 overexpression plants, suggesting that DHAR3 is downstream of HRG1. That the root meristem length and cell number were significantly reduced in hrg1-1 and hrg2-1 plants upon H2O2 treatment compared to that of HRG1 overexpression plants also approves the idea that HRGs function in H2O2 removal. Further evolutionary analysis indicates that this is a dicotyledon-specific pathway responsive to H2O2. Together, this work reveals HRG1/2 as novel H2O2 responders involved in ROS scavenging to ensure embryonic root meristem activity. These findings provide valuable clues for the of H2O2 signaling and root meristem regulation.

Characterization of a salt-resistant fibrinolytic protease of Bacillus licheniformis HJ4 isolated from Hwangseokae jeotgal, a traditional Korean fermented seafood.

Bacillus licheniformis HJ4 showing strong fibrinolytic activity was isolated from Hwangseokae jeotgal. aprEHJ4, a major fibrinolytic gene, was cloned by PCR, and an ORF consisting of 379 amino acids was located. The mature enzyme was expected to be 27 kDa in size after processing, but a 24-kDa protein was observed by SDS-PAGE and fibrin zymography, indicating additional processing. RT-qPCR showed that expression level of aprEHJ4 in culture with 0% salt (control) was the highest followed by culture with 8% salt (89.7% of control) and 5% salt (74.2%) at 84 h. The expression level in culture with 15% salt was 46.9%. The results matched with the fibrinolytic activity measurements of cultures and indicated that AprEHJ4 maintained significant activity in the presence of salt up to 15% (w/v). AprEHJ4 was overproduced in Escherichia coli, and mature 27 kDa protein was purified after in vitro renaturation. The optimum pH and temperature of AprEHJ4 were pH 8 and 40 ℃, respectively.

Increase of a Fibrinolytic Enzyme Production through Promoter Replacement of aprE3-5 from Bacillus subtilis CH3-5.

Bacillus subtilis CH3-5 isolated from cheonggukjang secretes a 28 kDa protease with a strong fibrinolytic activity. Its gene, aprE3-5, was cloned and expressed in a heterologous host (Jeong et al., 2007). In this study, the promoter of aprE3-5 was replaced with other stronger promoters (Pcry3A, P10, PSG1, PsrfA) of Bacillus spp. using PCR. The constructed chimeric genes were cloned into pHY300PLK vector, and then introduced into B. subtilis WB600. The P10 promoter conferred the highest fibrinolytic activity, i.e., 1.7-fold higher than that conferred by the original promoter. Overproduction of the 28 kDa protease was confirmed using SDS-PAGE and fibrin zymography. RT-qPCR analysis showed that aprE3-5 expression was 2.0-fold higher with the P10 promoter than with the original promoter. Change of the initiation codon from GTG to ATG further increased the fibrinolytic activity. The highest aprE3-5 expression was observed when two copies of the P10 promoter were placed in tandem upstream of the ATG initiation codon. The construct with P10 promoter and ATG and the construct with two copies of P10 promoter in tandem and ATG exhibited 117% and 148% higher fibrinolytic activity, respectively, than that exhibited by the construct containing P10 promoter and GTG. These results confirmed that significant overproduction of a fibrinolytic enzyme can be achieved by suitable promoter modification, and this approach may have applications in the industrial production of AprE3-5 and related fibrinolytic enzymes.

Cloning of a Novel vpr Gene Encoding a Minor Fibrinolytic Enzyme from Bacillus subtilis SJ4 and the Properties of Vpr.

We have previously characterized AprESJ4, the major fibrinolytic enzyme from Bacillus subtilis SJ4 (Yao et al., 2019). During that study, we observed a 68 kDa protein with fibrinolytic activity. In this study, we cloned the gene (vprSJ4) encoding the 68 kDa protein, a mature Vpr and minor protease secreted by Bacillus species. vprSJ4 encodes a preproenzyme consisting of 810 amino acids (aa) including signal sequence (28 aa) and prosequence (132 aa). The mature enzyme (650 aa) has a predicted molecular weight of 68,467.35. Unlike Vprs from other B. subtilis strains, VprSJ4 has 4 additional amino acids (DEFA) at the C-terminus. vprSJ4 was overexpressed in Escherichia coli. PreproVprSJ4 was localized in inclusion bodies, and subjected to in vitro renaturation and purification by an affinity column. SDS-PAGE and western blot showed that autoprocessing of preproVprSJ4 occurred and 68 kDa and smaller proteins were produced. The optimum pH and temperature of the recombinant VprSJ4 were pH 7.0 and 40°C, respectively. Kinetic parameters of recombinant VprSJ4 were measured by using an artificial substrate, N-succinyl-ala-ala-pro-phe-p-nitroanilide. Coexpression of vprSJ4 and aprESJ4 using pHY300PLK increased the fibrinolytic activity a further 117% when compared with aprESJ4 single expression using the same vector in B. subtilis WB600.

Na-Based monolayer photocatalysts with an extremely high intrinsic electric-field for water splitting.

A strong built-in electric field, high carrier mobility and a wide range of optical absorption values are the key parameters for photocatalysts used in water splitting. The design and preparation of photocatalysts possessing simultaneously these characteristics have always been the main tasks in the water splitting field. Here, we report a new family of 2D Na-based photocatalysts, NaAB2 (A = Al, Ga, In; B = S, Se, Te) monolayers, which may achieve this goal. First-principles calculations show that most of the NaAB2 monolayers are semiconductors with a suitable direct band gap ranging from visible to near-infrared light, exhibiting good optical absorption. The electron mobilities of the NaAB2 monolayers are up to 103 cm2 V-1 s-1, meaning the rapid migration of electrons can promote photocatalytic overall water splitting. Importantly, the electrostatic potential differences between the top surface and the bottom surface are larger than 1.23 eV for all the studied NaAB2 monolayers, meaning a high intrinsic built-in electric field that is present in these Na-based photocatalysts can promote the overall water splitting irrespective of their band gaps and band edges. Our studies show that the NaAB2 monolayers may be ideal photocatalysts for use in water splitting and may initiate a new round of experimental studies.

Multidimensional Vibration Suppression Method with Piezoelectric Control for Wind Tunnel Models.

In wind tunnel tests, the low-frequency and large-amplitude vibration of the cantilever sting result in poor test data in pitch plane and yaw plane, more seriously, even threatens the safety of wind tunnel tests. To ensure the test data quality, a multidimensional vibration suppression method is proposed to withstand the vibration from any direction, which is based on a system with stackable piezoelectric actuators and velocity feedback employing accelerometers. Firstly, the motion equation of the cantilever sting system is obtained by Hamilton's principle with the assumed mode method. Then, the multidimensional active control mechanism is qualitatively analyzed and a negative velocity feedback control algorithm combined with a root mean square (RMS) evaluation method is introduced to realize active mass and active damping effect, meanwhile, a weight modification method is performed to determine the sequence number of the stacked piezoelectric actuators and the weight of control voltages in real time. Finally, a multidimensional vibration suppression system was established and verification experiments were carried out in lab and a transonic wind tunnel. The results of lab experiments indicate that the damping ratio of the system is improved more than 4.3 times and the spectrum analyses show reductions of more than 23 dB. In addition, wind tunnel test results have shown that for the working conditions (α = -4~10° with γ = 0° or α = -4~10° with γ = 45°) respectively at 0.6 Ma and 0.7 Ma, the remainder vibration is less than 1.53 g, which proves that the multidimensional vibration suppression method has the ability to resist vibration from any direction to ensure the smooth process of wind tunnel tests.

Purification and Characterization of a Bacteriocin, BacBS2, Produced by Bacillus velezensis BS2 Isolated from Meongge Jeotgal.

Bacillus velezensis BS2 was isolated from meongge (common sea squirt) jeotgal, a Korean fermented seafood, and produces a bacteriocin, BacBS2, which strongly inhibits Listeria monocytogenes and Bacillus cereus. BacBS2 was partially purified by Q-Sepharose column chromatography after ammonium sulfate precipitation of the culture supernatant, then further purified by Sephadex G-50 column chromatography. Partially purified BacBS2 was estimated to be 6.5 kDa in size by Tricine-SDS PAGE and activity detection by gel-overlay. Enzyme treatment and FT-IR spectrum of partially purified BacBS2 confirmed its proteinaceous nature. BacBS2 was fully stable at pH 4-9, and half of activity was retained at pH 1-3. Full activity was retained after exposure to 80°C for 15 min, but half of the activity was retained upon exposure to 90°C for 15 min or 100°C for 10 min. BacBS2 inhibited L. monocytogenes by bactericidal mode of action. B. velezensis BS2 and its BacBS2 seem useful as biopreservatives for fermented foods such as jeotgal.

Physicochemical Properties and Bacterial Communities of Meongge (Halocynthia roretzi) Jeotgal Prepared with 3 Different Types of Salts.

Three types of meongge (Halocynthia roretzi) jeotgal (MJ) were prepared with 3 different types of salts (12%, w/v): purified salt (PS), solar salt aged for 3 years (SS), and bamboo salt that had been recrystalized 3 times (BS). One set of MJ was fermented with starters, Bacillus subtilis JS2 and Tetragenococcus halophilus BS1-37 (each 6 log CFU/g), and another set without starters for 42 days at 10°C. The LAB count of the SSMJ (non-starter) was highest at day 28 (2.30 log CFU/g). The pH of the PSMJ and SSMJ was 5.72-5.77 at day 0, and 5.40-5.50 at day 42. BSMJ showed higher pH and lower titratable acidities than other samples. Amino-type nitrogen (ANN) increased continuously, and SSMJ showed higher values than other samples from day 14. Bacterial species of non-starter MJ were examined by culture independent method. Clone libraries of 16S rRNA genes were constructed in Escherichia coli from total DNA from nonstarter MJ samples at day 0, 14, and 28. Thirty clones per each sample were randomly selected and DNA sequences were analyzed. Variovorax sp., uncultured bacterium, and Acidovorax sp. were the most dominant group at day 0, 14, and 28, respectively. Lactobacillus sakei and Streptococcus sp. were the next dominant group in SSMJ at day 28. A Streptococcus sp. was detected from PSMJ at day 28. Sensory evaluation for MJ samples at day 28 showed that SSMJ got higher overall acceptability scores. These results showed that solar salt can cause desirable changes in the microbial community of fermented foods, thereby positively affecting their overall quality.

Characterization of a Fibrinolytic Enzyme Secreted by Bacillus velezensis BS2 Isolated from Sea Squirt Jeotgal.

Bacillus sp. BS2 showing strong fibrinolytic activity was isolated from sea squirt (munggae) jeotgal, a traditional Korean fermented seafood. BS2 was identified as B. velezensis by molecular biological methods.B. velezensis BS2 grows well at 15% NaCl and at 10oC. When B. velezensis BS2 was cultivated in TSB broth for 96 h at 37°C, the culture showed the highest fibrinolytic activity (131.15 mU/µl) at 96 h. Three bands of 27, 35 and 60 kDa were observed from culture supernatant by SDS-PAGE, and fibrin zymography showed that the major fibrinolytic protein was the 27 kDa band. The gene (aprEBS2) encoding the major fibrinolytic protein was cloned, and overexpressed in heterologous hosts, B. subtilis WB600 and E. coli BL21 (DE3). B. subtilis transformant showed 1.5-fold higher fibrinolytic activity than B. velezensis BS2. Overproduced AprEBS2 in E. coli was purified by affinity chromatography. The optimum pH and temperature were pH 8.0 and 37°C, respectively. Km and Vmax were 0.15 mM and 39.68 µM/l/min, respectively, when N-succinyl-Ala-Ala-Pro-Phe-pNA was used as the substrate. AprEBS2 has strong α-fibrinogenase and moderate ß-fibrinogenase activity. Considering its high fibrinolytic activity, significant salt tolerance, and ability to grow at 10°C, B. velezensis BS2 can be used as a starter for jeotgal.

Effects of different types of salts on the growth of lactic acid bacteria and yeasts during kimchi fermentation.

Kimchi was prepared with different types of salts: purified salt (PS), solar salt aged for 1 year (SS1), aged for 3 years (SS3), and bamboo salt (BS). Kimchi inoculated with Leuconostoc mesenteroides P30 (starter kimchi), and control kimchi (non-starter kimchi) were prepared, and stored at - 1 °C for 20 weeks. Titratable acidity values increased slowly and reached 0.96-1.01% (pH 3.73-3.83) at 20 weeks. Proportions of coccus-type lactic acid bacteria (LAB) among total LAB were higher in SS kimchi than PS kimchi. Among non-starter kimchi, the proportions were 44.7, 41.6, 29.7, and 32.1% for SS3, SS1, BS, and PS kimchi, respectively, at 2 weeks, and 11.5, 12.8, 6.7, and 5.8%, respectively, at 20 weeks. SS kimchi had much less yeast counts than PS kimchi. Among starter kimchi, yeasts were detected from PS kimchi at 10 weeks but not detected until 18 weeks from SS1 and BS kimchi and 20 weeks from SS3 kimchi.

Properties of a fibrinolytic enzyme secreted by Bacillus subtilis JS2 isolated from saeu (small shrimp) jeotgal.

Bacillus species were screened to be used as starters for jeotgals, salted and fermented Korean sea foods. A strain, JS2, showing strong fibrinolytic activity was isolated from saeu (small shrimp) jeotgal, and identified as Bacillus subtilis. Bacillus subtilis JS2 grew well at 20% (w/v) NaCl concentration. SDS-PAGE of culture supernatant from JS2 showed 3 major bands of 27, 29, and 60 kDa in size. Fibrin zymography showed that the 27 kDa band was the major fibrinolytic protein. The gene, aprEJS2, was cloned and introduced into B. subtilis WB600 using pHY300PLK. A B. subtilis transformant harboring pHYJS2 showed higher fibrinolytic activity than B. subtilis JS2. aprEJS2 was overexpressed in Escherichia coli BL21 (DE3). The optimum pH and temperature for AprEJS2 were pH 8.0 and 40 °C, respectively. Km and Vmax values were determined. AprEJS2 has strong α-fibrinogenase activity and moderate ß-fibrinogenase activity.

Fascin induces melanoma tumorigenesis and stemness through regulating the Hippo pathway.

BACKGROUND: Fascin is a F-actin bundling protein and its overexpression is correlated with poor prognosis and increases metastatic potential in a number of cancers. But underlying function and mechanism of fascin on tumorigenesis in melanoma remain elusive. METHODS: The melanoma cell lines WM793 and WM39 were employed for the soft agar and sphere formation assay. Quantitative RT-PCR and Western blot were performed for identifying the gene expression at mRNA and protein levels, respectively. Co-IP and in vitro GST pulldown experiments were used to test the interaction between fascin and MST2. RESULTS: Fascin regulates tumorigenesis and cancer cell stemness in melanoma through inhibition of the Hippo pathway kinase MST2 and the activation of transcription factor TAZ. Our data showed that fascin interacts with the kinase domain of MST2 to inhibit its homodimer formation and kinase activity. Depletion of fascin led to increase of p-LATS level and decrease of TAZ, but not YAP. We also demonstrated that fascin regulates melanoma tumorigenesis independent of its actin-bundling activity. CONCLUSIONS: Fascin is a new regulator of the MST2-LATS-TAZ pathway and plays a critical role in melanoma tumorigenesis. Inhibition of fascin reduces melanoma tumorigenesis and stemness, and thus fascin could be a potential therapeutic target for this malignancy.