Application of ultrasensitive assay for SARS-CoV-2 antigen in nasopharynx in the management of COVID-19 patients with comorbidities during the peak of 2022 Shanghai epidemics in a tertiary hospital.

OBJECTIVES: Various comorbidities associated with COVID-19 add up in severity of the disease and obviously prolonged the time for viral clearance. This study investigated a novel ultrasensitive MAGLUMI® SARS-CoV-2 Ag chemiluminescent immunoassay assay (MAG-CLIA) for diagnosis and monitoring the infectivity of COVID-19 patients with comorbid conditions during the pandemic of 2022 Shanghai. METHODS: Analytical performances of the MAG-CLIA were evaluated, including precision, limit of quantitation, linearity and specificity. Nasopharyngeal specimens from 232 hospitalized patients who were SARS-CoV-2 RT-qPCR positive and from 477 healthy donors were included. The longitudinal studies were performed by monitoring antigen concentrations alongside with RT-qPCR results in 14 COVID-19 comorbid participants for up to 22 days. The critical antigen concentration in determining virus infectivity was evaluated at the reference cycle threshold (Ct) of 35. RESULTS: COVID-19 patients were well-identified using an optimal threshold of 0.64 ng/L antigen concentration, with sensitivity and specificity of 95.7% (95% CI: 92.2-97.9%) and 98.3% (95% CI: 96.7-99.3%), respectively, while the Wondfo LFT exhibited those of 34.9% (95% CI: 28.8-41.4%) and 100% (95% CI: 99.23-100%), respectively. The sensitivity of MAG-CLIA remained 91.46% (95% CI: 83.14-95.8%) for the samples with Ct values between 35 and 40. Close dynamic consistence was observed between MAG-CLIA and viral load time series in the longitudinal studies. The critical value of 8.82 ng/L antigen showed adequate sensitivity and specificity in evaluating the infectivity of hospitalized convalescent patients with comorbidities. CONCLUSIONS: The MAG-CLIA SARS-CoV-2 Ag detection is an effective and alternative approach for rapid diagnosis and enables us to evaluate the infectivity of hospitalized convalescent patients with comorbidities.

More negative charges on roots enhanced manganese(II) uptake in leguminous and non-leguminous poaceae crops.

BACKGROUND: Manganese (Mn) is an essential micronutrient for plants, whereas excess Mn(II) in soils leads to its toxicity to crops. Mn(II) is adsorbed onto plant roots from soil solution and then absorbed by plants. Root charge characteristics should affect Mn(II) toxicity to crops and Mn(II) uptake by the roots of the crops. However, the differences in the effects of root surface charge on the uptake of Mn(II) among various crop species are not well understood. RESULTS: The roots of nine legumes and six non-legume poaceae were obtained by hydroponics and the streaming potential method and spectroscopic analysis were used to measure the zeta potentials and functional groups on the roots, respectively. The results indicate that the exchangeable Mn(II) adsorbed by plant roots was significantly positively correlated with the Mn(II) accumulated in plant shoots. Legume roots carried more negative charges and functional groups than non-legume poaceae roots, which was responsible for the larger amounts of exchangeable Mn(II) on legume roots in 2 h and the Mn(II) accumulated in their shoots in 48 h. Coexisting cations, such as Ca2+ and Mg2+ , were most effective in decreasing Mn(II) taken up by roots and accumulated in shoots than K+ and Na+ . This was because Ca2+ and Mg2+ could compete with Mn(II) for active sites on plant roots more strongly compared to K+ and Na+ . CONCLUSION: The root surface charge and functional groups are two important factors influencing Mn(II) uptake by roots and accumulation in plant shoots. © 2023 Society of Chemical Industry.

Resistance of Acta2R149C/+ mice to aortic disease is associated with defective release of mutant smooth muscle α-actin from the chaperonin-containing TCP1 folding complex.

Pathogenic variants of the gene for smooth muscle α-actin (ACTA2), which encodes smooth muscle (SM) α-actin, predispose to heritable thoracic aortic disease. The ACTA2 variant p.Arg149Cys (R149C) is the most common alteration; however, only 60% of carriers have a dissection or undergo repair of an aneurysm by 70 years of age. A mouse model of ACTA2 p.Arg149Cys was generated using CRISPR/Cas9 technology to determine the etiology of reduced penetrance. Acta2R149C/+ mice had significantly decreased aortic contraction compared with WT mice but did not form aortic aneurysms or dissections when followed to 24 months, even when hypertension was induced. In vitro motility assays found decreased interaction of mutant SM α-actin filaments with SM myosin. Polymerization studies using total internal reflection fluorescence microscopy showed enhanced nucleation of mutant SM α-actin by formin, which correlated with disorganized and reduced SM α-actin filaments in Acta2R149C/+ smooth muscle cells (SMCs). However, the most prominent molecular defect was the increased retention of mutant SM α-actin in the chaperonin-containing t-complex polypeptide folding complex, which was associated with reduced levels of mutant compared with WT SM α-actin in Acta2R149C/+ SMCs. These data indicate that Acta2R149C/+ mice do not develop thoracic aortic disease despite decreased contraction of aortic segments and disrupted SM α-actin filament formation and function in Acta2R149C/+ SMCs. Enhanced binding of mutant SM α-actin to chaperonin-containing t-complex polypeptide decreases the mutant actin versus WT monomer levels in Acta2R149C/+ SMCs, thus minimizing the effect of the mutation on SMC function and potentially preventing aortic disease in the Acta2R149C/+ mice.

Unusual dynamics of the divergent malaria parasite PfAct1 actin filament.

Gliding motility and host cell invasion by the apicomplexan parasite Plasmodium falciparum (Pf), the causative agent of malaria, is powered by a macromolecular complex called the glideosome that lies between the parasite plasma membrane and the inner membrane complex. The glideosome core consists of a single-headed class XIV myosin PfMyoA and a divergent actin PfAct1. Here we use total internal reflection fluorescence microscopy to visualize growth of individual unstabilized PfAct1 filaments as a function of time, an approach not previously used with this actin isoform. Although PfAct1 was thought to be incapable of forming long filaments, filaments grew as long as 30 µm. Polymerization occurs via a nucleation-elongation mechanism, but with an ∼4 µM critical concentration, an order-of-magnitude higher than for skeletal actin. Protomers disassembled from both the barbed and pointed ends of the actin filament with similar fast kinetics of 10 to 15 subunits/s. Rapid treadmilling, where the barbed end of the filament grows and the pointed end shrinks while maintaining an approximately constant filament length, was visualized near the critical concentration. Once ATP has been hydrolyzed to ADP, the filament becomes very unstable, resulting in total dissolution in <40 min. Dynamics at the filament ends are suppressed in the presence of inorganic phosphate or more efficiently by BeFX A chimeric PfAct1 with a mammalian actin D-loop forms a more stable filament. These unusual dynamic properties distinguish PfAct1 from more canonical actins, and likely contribute to the difficultly in visualizing PfAct1 filaments in the parasite.

Effects of pH variations caused by redox reactions and pH buffering capacity on Cd(II) speciation in paddy soils during submerging/draining alternation.

Incubation experiments were conducted to investigate the influencing factors of pH variation in different paddy soils during submerging/draining alternation and the relationship between pH buffering capacity (pHBC) and Cd speciation in ten paddy soils developed from different parent materials (including 8 acid paddy soils and 2 alkaline paddy soils). The soil pHBC and the changes in soil pH, Eh, Fe2+, Mn2+, SO42- and Cd speciation were determined. The results showed that there was a significant positive correlation between cation exchange capacity (CEC) and pHBC of these paddy soils, indicating that soil CEC is a key factor affecting the pHBC of paddy soils. The contribution of Fe(III) oxide reduction to H+ consumption is far greater than the reduction of Mn(IV)/Mn(III) oxides and SO42- during the submerging. For example, the contribution of the reduction of manganese oxides, SO42- and iron oxides to H+ consumption in the paddy soils from Anthrosol at 15 d submerging was 1.2%, 11.6% and 87.2%, respectively. This confirms that the reduction of Fe(III) oxides plays a leading role in increasing soil pH. Importantly, we noticed that during submerging, soil pH was increased and resulted in the content of available Cd in soils being reduced. This was due to the transformation of Cd to less active forms. Also, there was a significant positive correlation between the change rate of available Cd, the percentage of acid extractable Cd and pH variation. This suggests that the variation in soil pH was responsible for the transformation of Cd speciation. In addition, the change rate of available Cd and the percentage of acid extractable Cd concentration were significantly negatively correlated with soil pHBC. The soil with higher pHBC experienced less pH change, and thus the change rate of available Cd and the percentage of acid extractable Cd concentration were less for the soil. The results of this study can provide a basis for the remediation of Cd-contaminated acidic paddy soils.

Molecular Insights into the Effect of Nitrogen Bubbles on the Formation of Tetrahydrofuran Hydrates.

In this work, a molecular dynamics simulation was conducted to study the microscopic mechanism of how nitrogen bubbles affect the formation of THF hydrates at the molecular level. The results obtained reveal that the nitrogen bubble can promote the formation of THF hydrates. In the system with a nitrogen bubble, more THF-filled cages were generated, and the crystal structure was more orderly. The promotion of nitrogen bubbles on hydrate crystallization comes from the dissolution of nitrogen molecules. Some of dissolved nitrogen molecules can be enclosed in small hydrate cages near the nitrogen bubble, which can serve as stable sites for hydrate crystal growth, resulting in the fact that THF-filled cages connected with N2-filled cages are much more stable and have a long lifetime. The results in this work can help to understand the promotion effect of micro- and nano-air bubbles on the crystallization of THF hydrates.

Geochemical Characteristics and Significance of Organic Matter in Hydrate-Bearing Sediments from Shenhu Area, South China Sea.

Rock-Eval pyrolysis and the biomarker composition of organic matter were systematically studied in hydrate-bearing sediments from the Shenhu area, South China Sea. The n-alkane distribution patterns revealed that the organic matter in the sediments appeared to originate from mixed sources of marine autochthonous input, terrestrial higher plants, and ancient reworked organic matter. The low total organic carbon contents (average < 0.5%) and the low hydrogen index (HI, <80 mg HC/g TOC) suggested the poor hydrocarbon-generation potential of the deposited organic matter at a surrounding temperature of <20 °C in unconsolidated sediments. The abnormally high production index and the fossil-originated unresolved complex mixture (UCM) accompanied by sterane and hopane of high maturity indicated the contribution of deep hydrocarbon reservoirs. Preliminary oil-to-source correlation for the extracts implied that the allochthonous hydrocarbons in the W01B and W02B sediments might have originated from the terrestrial source rocks of mature Enping and Wenchang formations, while those of W03B seem to be derived from more reduced and immature marine source rocks such as the Zhuhai formation. The results of the organic extracts supported the previous identification of source rocks based on the isotopic composition of C2+ hydrate-bound gases. The biomarker of methanogens, squalane, was recognized in the sediments of this study, possibly suggesting the generation of secondary microbial gases which are coupled with the biodegradation of the deep allochthonous hydrocarbons.

Effect of root surface charge on the absorption and accumulation of Cu(II) by different japonica and indica rice varieties under acidic conditions.

Excessive amounts of copper (Cu) in soils causes toxic effects on plants. In this study, 58 rice cultivars were classified into tolerant, moderately tolerant, and susceptible types for Cu(II) toxicity based on 50% germination (LC50). Nine japonica rice varieties (three each from the tolerant, moderately tolerant, and susceptible groups) and six indica rice varieties (three from the moderately tolerant and susceptible groups) were selected for the hydroponics experiments. In the short-term adsorption experiment, Cu(II) adsorbed on rice roots was differentiated into exchangeable, complexed, and precipitated forms. Similarly, it was done for long-term culture. Absorption of Cu(II) by rice roots and shoots was also measured. The results indicated that adsorbed Cu(II) mainly existed as complexed and exchangeable forms on rice roots in the short-term adsorption experiment, and the exchangeable and complexed Cu(II) levels were greater for indica rice than for japonica rice due to the larger negative charge on the indica rice roots. The adsorbed Cu(II) mainly existed as a complexed form in the long-term culture experiment, and the exchangeable Cu(II) level was much lower than that in the short-term adsorption experiment due to the absorption of Cu(II) by rice plants. The indica varieties absorbed more Cu(II) than the japonica varieties. Furthermore, the absorption and accumulation of Cu(II) by the susceptible varieties were greater than by the tolerant and moderately tolerant varieties for both the japonica and indica rice. The absorption and accumulation of Cu(II) in rice roots were much greater than in the shoots. Chlorophyll content, and the lengths and dry matter weights of the rice roots and shoots decreased with increasing Cu(II) concentration. The Cu(II) showed greater toxicity toward indica varieties than japonica varieties, and the greater negative charge on indica roots was one of reasons for the greater exchangeable Cu(II) on the roots, the increase in Cu(II) toxicity, and the higher uptake of Cu(II) by indica rice varieties compared to japonica rice varieties.

Effects of surface charge and chemical forms of manganese(II) on rice roots on manganese absorption by different rice varieties.

The roots of 4 japonica, 4 indica, and 7 hybrid rice varieties were obtained by hydroponic experiment and used to explore the relationship between charge characteristics and exchangeable manganese(II) (Mn(II)) on rice roots and Mn(II) absorption in roots and shoots of the rice. Results indicated Mn(II) adsorbed on rice roots mainly existed as exchangeable Mn(II) after 2 h. The roots of indica and hybrid rice carried more negative charges than the roots of japonica rice. Accordingly, this led to more exchangeable Mn(II) to be adsorbed on roots of indica and hybrid rice after 2 h and more Mn(II) absorbed in the roots of the same varieties after 48 h. However, this was contrary to the result of Mn(II) absorption in rice shoots after 48 h. Coexisting cations of K+, Na+, Ca2+, and Mg2+ reduced the exchangeable Mn(II) on rice roots through their competition with Mn(II) for sorption sites on rice roots, which led to the decrease in Mn(II) absorption in rice roots and shoots. Ca2+ and Mg2+ showed a greater decrease in the Mn(II) absorbed in roots and shoots than K+ and Na+. The reduction of Mn(II) absorption in the roots of indica rice and hybrid rice induced by Ca2+ and Mg2+ was more than that of japonica rice. This was attributed to more negative charges on the roots of the former than the latter. Therefore, the absorption of Mn(II) by rice roots was determined by surface charge properties and exchangeable Mn(II) on the rice roots. The results suggested that Ca2+ and Mg2+ have potential to alleviate Mn(II) toxicity to rice.

Comparing ameliorative effects of biomass ash and alkaline slag on an acidic Ultisol under artificial Masson pine: A field experiment.

Forest soil acidification caused by acid deposition is a serious threat to the forest ecosystem. To investigate the liming effects of biomass ash (BA) and alkaline slag (AS) on the acidic topsoil and subsoil, a three-year field experiment under artificial Masson pine was conducted at Langxi, Anhui province in Southern China. The surface application of BA and AS significantly increased the soil pH, and thus decreased exchangeable acidity and active Al in the topsoil. Soil exchangeable Ca2+ and Mg2+ in topsoil were significantly increased by the surface application of BA and AS, while an increase in soil exchangeable K+ was only observed in BA treatments. The soil acidity and active Al in subsoil were decreased by the surface application of AS. Compared with the control, soluble monomeric and exchangeable Al in the subsoil was decreased by 38.0% and 29.4% after 3 years of AS surface application. There was a minimal effect on soluble monomeric and exchangeable Al after the application of BA. The soil exchangeable Ca2+ and Mg2+ in the subsoil increased respectively by 54% and 141% after surface application of 10 t ha-1 AS. The decrease of soil active Al and increase of base cations in subsoil were mainly attributed to the high migration capacity of base cations in AS. In conclusion, the effect of surface application of AS was superior to BA in ameliorating soil acidity and alleviating soil Al toxicity in the subsoil of this Ultisol.

MAP7 regulates organelle transport by recruiting kinesin-1 to microtubules.

Microtubule-associated proteins (MAPs) regulate microtubule polymerization, dynamics, and organization. In addition, MAPs alter the motility of kinesin and dynein to control trafficking along microtubules. MAP7 (ensconsin, E-MAP-115) is a ubiquitous MAP that organizes the microtubule cytoskeleton in mitosis and neuronal branching. MAP7 also recruits kinesin-1 to microtubules. To understand how the activation of kinesin-1 by MAP7 regulates the motility of organelles transported by ensembles of kinesin and dynein, we isolated organelles and reconstituted their motility in vitro In the absence of MAP7, isolated phagosomes exhibit approximately equal fractions of plus- and minus-end-directed motility along microtubules. MAP7 causes a pronounced shift in motility; phagosomes move toward the plus-end ∼80% of the time, and kinesin teams generate more force. To dissect MAP7-mediated regulation of kinesin-driven transport, we examined its effects on the motility and force generation of single and teams of full-length kinesin-1 motors. We find that MAP7 does not alter the force exerted by a single kinesin-1 motor, but instead increases its binding rate to the microtubule. For ensembles of kinesin, a greater number of kinesin motors are simultaneously engaged and generating force to preferentially target organelles toward the microtubule plus-end.

On the phase fading effect in the dual-pulse heterodyne demodulated distributed acoustic sensing system.

Understanding signal fading effect is essential for the application of Rayleigh-scattering-based distributed acoustic fibre sensors (DASs) due to the nature of coherent beam interference within the pulse length. Statistical properties for the intensity of the Rayleigh backscattered light (i.e. intensity fading) and its impact on the sensitivity of DAS systems have been intensely studied over the last decades. Here we for the first time establish an analytical model for the phase signal retrieved from the dual-pulse heterodyne demodulated DAS system, which can be exploited to investigate the phase fading effect in this system. The developed model reveals that the phase fading phenomenon mainly originates from the randomness in the phase retardant of the Rayleigh scatters. The quantitatively resolved statistical features of the phase fading is confirmed by experimental results. Based on the analytical model, a noise figure is defined to characterize the global fading-induced noise level via taking into account contributions from all channels along the sensing fiber. The model also reproduces the anti-correlation relation between the power spectrum density of retrieved phase at the heterodyne frequency and the phase fading noise level. Following the analysis and the definition of the noise figure, an optimized real-time weighted-channel stack algorithm is developed to efficiently suppress the fading noise. Experimental results show that the algorithm can achieve a maximum noise figure reduction of 15.8 dB without increasing the system complexity.

Method for initially selecting Al-tolerant rice varieties based on the charge characteristics of their roots.

To explore the relationship between charge characteristics of rice roots and aluminum (Al) tolerance of rice, roots of 47 different rice genotypes were obtained by hydroponic experiment. The zeta potentials of roots were determined by streaming potential method, and the Al tolerance and the functional groups of rice were measured by relative root elongation and infrared spectroscopy (ATR-FTIR), respectively. The exchangeable, complexed and precipitated Al(III) sorbed on the root surface of rice was extracted with 1 mol L-1 KNO3, 0.05 mol L-1 EDTA-2Na and 0.01 mol L-1 HCl, respectively. There was a significant correlation between the zeta potentials and the relative elongation of rice roots, indicating that the zeta potentials of rice roots could be used to characterize rice tolerance to Al toxicity. Twelve Al-tolerant rice varieties, 25 medium Al-tolerant rice varieties, and 10 Al-sensitive rice varieties were obtained. The Al-tolerant rice varieties sorbed less complexed Al(III) and total Al(III) because there was lower negative charge on their roots compared to less tolerant genotypes. A correlation analysis showed that there were significant negative correlations between the zeta potential, relative root elongation, and the total Al(III) sorption capacity of the roots, which further confirmed the reliability of using the root zeta potential to characterize rice tolerance to Al toxicity. The results of this paper provide a new method for screening Al-tolerant rice varieties.

Phytotoxicity of Cu2+ and Cd2+ to the roots of four different wheat cultivars as related to charge properties and chemical forms of the metals on whole plant roots.

The relationship between the chemical forms of Cu2+ and Cd2+ adsorbed on the roots of different wheat cultivars and their phytotoxic effects on the plants were investigated. The wheat varieties Dunmaiwang (DMW), Tekang 6 (TK6), Zhongmai895 (ZM895), and Chaojixiaomai (AK68) were used. The zeta potentials of wheat roots, measured by the streaming potential method, were used to characterize root charge properties. Results indicated that the changes in zeta potential at pH 4.01-6.61 were 14.7, 15.53, 13.01, and 12.06 mV for ZM895, AK68, DMW, and TK6, respectively. The negative charge and functional groups on ZM895 and AK68 roots were greater than on DMW and TK6 roots, which led to more exchangeable and complexed Cu2+ and Cd2+ on ZM895 and AK68 roots and increased Cu2+ and Cd2+ toxicity compared to DMW and TK6. Coexisting cations, such as Ca2+, Mg2+, K+, and NH4+, alleviated Cu2+ and Cd2+ toxicity to wheat roots through competition for adsorption sites on the roots, which decreased exchangeable and complexed Cu2+ and Cd2+ on wheat roots. The Ca2+ and Mg2+ were most effective in alleviating heavy metal toxicity and they decreased exchangeable Cu2+ on AK68 roots by 39.14% and 47.82%, and exchangeable Cd2+ by 8.51% and 28.23%, respectively.

Performance Improvement of Dual-Pulse Heterodyne Distributed Acoustic Sensor for Sound Detection.

Phase fading is fatal to the performance of distributed acoustic sensors (DASs) influencing its capability of distributed measurement as well as its noise level. Here, we report the experimental observation of a strong negative correlation between the relative power spectrum density (PSD) at the heterodyne frequency and the noise floor of the detected phase for the heterodyne demodulated distributed acoustic sensor (HD-DAS) system. We further propose a weighted-channel stack algorithm (WCSA) to alleviate the phase fading noise via an enhancement of the relative PSD at the heterodyne frequency. Experimental results show that the phase noise of the demodulated signal can be suppressed by 13.7 dB under optimal condition. As a potential application, we exploited the improved HD-DAS system to retrieve a piece of music lasted for 205 s, demonstrating the reliability of detecting wideband sound signal without distortion.

Reconstitution of the core of the malaria parasite glideosome with recombinant Plasmodium class XIV myosin A and Plasmodium actin.

Motility of the apicomplexan malaria parasite Plasmodium falciparum is enabled by a multiprotein glideosome complex, whose core is the class XIV myosin motor, PfMyoA, and a divergent Plasmodium actin (PfAct1). Parasite motility is necessary for host-cell invasion and virulence, but studying its molecular basis has been hampered by unavailability of sufficient amounts of PfMyoA. Here, we expressed milligram quantities of functional full-length PfMyoA with the baculovirus/Sf9 cell expression system, which required a UCS (UNC-45/CRO1/She4p) family myosin chaperone from Plasmodium spp. In addition to the known light chain myosin tail interacting protein (MTIP), we identified an essential light chain (PfELC) that co-purified with PfMyoA isolated from parasite lysates. The speed at which PfMyoA moved actin was fastest with both light chains bound, consistent with the light chain-binding domain acting as a lever arm to amplify nucleotide-dependent motions in the motor domain. Surprisingly, PfELC binding to the heavy chain required that MTIP also be bound to the heavy chain, unlike MTIP that bound the heavy chain independently of PfELC. Neither the presence of calcium nor deletion of the MTIP N-terminal extension changed the speed of actin movement. Of note, PfMyoA moved filaments formed from Sf9 cell-expressed PfAct1 at the same speed as skeletal muscle actin. Duty ratio estimates suggested that as few as nine motors can power actin movement at maximal speed, a feature that may be necessitated by the dynamic nature of Plasmodium actin filaments in the parasite. In summary, we have reconstituted the essential core of the glideosome, enabling drug targeting of both of its core components to inhibit parasite invasion.

Vascular disease-causing mutation R258C in ACTA2 disrupts actin dynamics and interaction with myosin.

Point mutations in vascular smooth muscle α-actin (SM α-actin), encoded by the gene ACTA2, are the most prevalent cause of familial thoracic aortic aneurysms and dissections (TAAD). Here, we provide the first molecular characterization, to our knowledge, of the effect of the R258C mutation in SM α-actin, expressed with the baculovirus system. Smooth muscles are unique in that force generation requires both interaction of stable actin filaments with myosin and polymerization of actin in the subcortical region. Both aspects of R258C function therefore need investigation. Total internal reflection fluorescence (TIRF) microscopy was used to quantify the growth of single actin filaments as a function of time. R258C filaments are less stable than WT and more susceptible to severing by cofilin. Smooth muscle tropomyosin offers little protection from cofilin cleavage, unlike its effect on WT actin. Unexpectedly, profilin binds tighter to the R258C monomer, which will increase the pool of globular actin (G-actin). In an in vitro motility assay, smooth muscle myosin moves R258C filaments more slowly than WT, and the slowing is exacerbated by smooth muscle tropomyosin. Under loaded conditions, small ensembles of myosin are unable to produce force on R258C actin-tropomyosin filaments, suggesting that tropomyosin occupies an inhibitory position on actin. Many of the observed defects cannot be explained by a direct interaction with the mutated residue, and thus the mutation allosterically affects multiple regions of the monomer. Our results align with the hypothesis that defective contractile function contributes to the pathogenesis of TAAD.

Severe Molecular Defects Exhibited by the R179H Mutation in Human Vascular Smooth Muscle α-Actin.

Mutations in vascular smooth muscle α-actin (SM α-actin), encoded by ACTA2, are the most common cause of familial thoracic aortic aneurysms that lead to dissection (TAAD). The R179H mutation has a poor patient prognosis and is unique in causing multisystemic smooth muscle dysfunction (Milewicz, D. M., Østergaard, J. R., Ala-Kokko, L. M., Khan, N., Grange, D. K., Mendoza-Londono, R., Bradley, T. J., Olney, A. H., Ades, L., Maher, J. F., Guo, D., Buja, L. M., Kim, D., Hyland, J. C., and Regalado, E. S. (2010) Am. J. Med. Genet. A 152A, 2437-2443). Here, we characterize this mutation in expressed human SM α-actin. R179H actin shows severe polymerization defects, with a 40-fold higher critical concentration for assembly than WT SM α-actin, driven by a high disassembly rate. The mutant filaments are more readily severed by cofilin. Both defects are attenuated by copolymerization with WT. The R179H monomer binds more tightly to profilin, and formin binding suppresses nucleation and slows polymerization rates. Linear filaments will thus not be readily formed, and cells expressing R179H actin will likely have increased levels of monomeric G-actin. The cotranscription factor myocardin-related transcription factor-A, which affects cellular phenotype, binds R179H actin with less cooperativity than WT actin. Smooth muscle myosin moves R179H filaments more slowly than WT, even when copolymerized with equimolar amounts of WT. The marked decrease in the ability to form filaments may contribute to the poor patient prognosis and explain why R179H disrupts even visceral smooth muscle cell function where the SM α-actin isoform is present in low amounts. The R179H mutation has the potential to affect actin structure and function in both the contractile domain of the cell and the more dynamic cytoskeletal pool of actin, both of which are required for contraction.

[Laser Raman Spectroscopy and Its Application in Gas Hydrate Studies].

Gas hydrates are important potential energy resources. Microstructural characterization of gas hydrate can provide information to study the mechanism of gas hydrate formation and to support the exploitation and application of gas hydrate technology. This article systemly introduces the basic principle of laser Raman spectroscopy and summarizes its application in gas hydrate studies. Based on Raman results, not only can the information about gas composition and structural type be deduced, but also the occupancies of large and small cages and even hydration number can be calculated from the relative intensities of Raman peaks. By using the in-situ analytical technology, laser Raman specstropy can be applied to characterize the formation and decomposition processes of gas hydrate at microscale, for example the enclathration and leaving of gas molecules into/from its cages, to monitor the changes in gas concentration and gas solubility during hydrate formation and decomposition, and to identify phase changes in the study system. Laser Raman in-situ analytical technology has also been used in determination of hydrate structure and understanding its changing process under the conditions of ultra high pressure. Deep-sea in-situ Raman spectrometer can be employed for the in-situ analysis of the structures of natural gas hydrate and their formation environment. Raman imaging technology can be applied to specify the characteristics of crystallization and gas distribution over hydrate surface. With the development of laser Raman technology and its combination with other instruments, it will become more powerful and play a more significant role in the microscopic study of gas hydrate.

The Therapeutic Potential of Hemp Seed Oil in D-Galactose-Induced Aging Rat Model Was Determined through the Combined Assessment of 1H NMR Metabolomics and 16S rRNA Gene Sequencing.

Aging is an irreversible process of natural degradation of bodily function. The increase in the aging population, as well as the rise in the incidence of aging-related diseases, poses one of the most pressing global challenges. Hemp seed oil, extracted from the seeds of hemp (Cannabis sativa L.), possesses significant nutritional and biological properties attributed to its unique composition of polyunsaturated fatty acids and various antioxidant compounds. However, there is limited knowledge regarding the anti-aging mechanism of hemp seed oil. This study aimed to evaluate the beneficial effects and potential mechanisms of hemp seed oil in a D-galactose (D-gal)-induced aging rat model through a combined analysis of metabolomics and 16S rRNA gene sequencing. Using nuclear magnetic resonance (NMR)-based metabolomics, significant alterations in serum and urine metabolic phenotypes were observed between the D-gal-induced aging rat model and the healthy control group. Eight and thirteen differentially expressed metabolites related to aging were identified in serum and urine, respectively. Treatment with hemp seed oil significantly restored four and ten potential biomarkers in serum and urine, respectively. The proposed pathways primarily included energy metabolism, amino acid metabolism, one-carbon metabolism, and lipid metabolism. Furthermore, 16S rRNA gene sequencing analysis revealed significant changes in the gut microbiota of aged rats. Compared to the model group, the hemp seed oil group exhibited significant alterations in the abundance of 21 bacterial taxa at the genus level. The results indicated that hemp seed oil suppressed the prevalence of pathogenic bacterial genera such as Streptococcus, Rothia, and Parabacteroides. Additionally, it facilitated the proliferation of the genera Lachnospirace_NK4B4_group and Lachnospirace_UCG_001, while also enhancing the relative abundance of the genus Butyricoccus; a producer of short-chain fatty acids (SCFAs). These findings provided new insights into the pathogenesis of aging and further supported the potential utility of hemp seed oil as an anti-aging therapeutic agent.