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Argonaute proteins (Agos) bind short nucleic acids as guides and are directed by them to recognize target complementary nucleic acids. Prokaryotic Agos (pAgos) are extremely diverse, with potential functions in microbial defense. The functions and mechanisms of a group of full-length yet inactive pAgos, long-B pAgos, remain enigmatic. Here, we show that most long-B pAgos constitute cell suicide systems together with their various associated proteins, including nucleases, Sir2-domain-containing proteins and trans-membrane proteins, respectively. Among them, the long-B pAgo-nuclease system utilizes an RNA-programmed and target-recognition-activated collateral DNA cleavage activity to sense invaders and mediate genomic DNA degradation. This kills the infected cells and results in depletion of the invader from the cell population. The data indicate that the long-B pAgo systems induce cell death with various effector proteins after recognition of invading nucleic acids, corresponding to an immune response via abortive infection.

SummaryArgonaute proteins (Agos) bind short nucleic acids as guides and are directed by them to recognize target complementary nucleic acids. Prokaryotic Agos (pAgos) are extremely diverse, with potential functions in microbial defense. The functions and mechanisms of a group of full-length yet inactive pAgos, long-B pAgos, remain enigmatic. Here, we show that most long-B pAgos constitute cell suicide systems together with their various associated proteins, including nucleases, Sir2-domain-containing proteins and trans-membrane proteins, respectively. Among them, the long-B pAgo-nuclease system utilizes an RNA-programmed and target-recognition-activated collateral DNA cleavage activity to sense invaders and kill the infected cells. This results in depletion of the invading plasmid from the cell population. Together, our data indicate that the long-B pAgo systems induce cell death with various effector proteins after recognition of invading nucleic acids, corresponding to an immune response via abortive infection.

CRISPR-Cas and prokaryotic Argonaute (pAgo) are nucleic acid (NA)-guided defense systems that protect prokaryotes against the invasion of mobile genetic elements. Previous studies established that they are directed by NA fragments (guides) to recognize invading complementary NA (targets), and that they cleave the targets to silence the invaders. Nevertheless, growing evidence indicates that many CRISPR-Cas and pAgo systems exploit the abortive infection (Abi) strategy to confer immunity. The CRISPR-Cas and pAgo Abi systems typically sense invaders using the NA recognition ability and activate various toxic effectors to kill the infected cells to prevent the invaders from spreading. This review summarizes the diverse mechanisms of these CRISPR-Cas and pAgo systems, and highlights their critical roles in the arms race between microbes and invaders.

Synthetic biology, a course with a sound theoretical system and a wide application range, plays a role in the cultivation of innovative talents in the field of bioengineering. To this end, we have set up a synthetic biology course in our university. First, according to the concept of imparting basic knowledge, highlighting innovative practice, and keeping up with cutting-edge progress, we assembled a high-level teaching team for synthetic biology. The team constantly adjusted and optimized the course contents and achieved a novel and reasonable course system. Second, we introduced frontier cases of synthetic biology reported in high-level journals, as well as breaking news in this field in classroom teaching, which enriched the teaching contents and aroused students' interest. Third, taking these cases as the breakthrough point, we guided students to in-depth discussions through the learning-centered teaching mode to improve students' abilities of critical thinking and theoretical innovation. In summary, the course has achieved good teaching outcomes and improved the cultivation of innovative talents. Therefore, we share our work with peer teachers, aiming to give new insights into the teaching reform of synthetic biology and other related courses.

Argonaute (Ago) proteins are widespread nucleic-acid-guided enzymes that recognize targets through complementary base pairing. Although, in eukaryotes, Agos are involved in RNA silencing, the functions of prokaryotic Agos (pAgos) remain largely unknown. In particular, a clade of truncated and catalytically inactive pAgos (short pAgos) lacks characterization. Here, we reveal that a short pAgo protein in the archaeon Sulfolobus islandicus, together with its two genetically associated proteins, Aga1 and Aga2, provide robust antiviral protection via abortive infection. Aga2 is a toxic transmembrane effector that binds anionic phospholipids via a basic pocket, resulting in membrane depolarization and cell killing. Ago and Aga1 form a stable complex that exhibits nucleic-acid-directed nucleic-acid-recognition ability and directly interacts with Aga2, pointing to an immune sensing mechanism. Together, our results highlight the cooperation between pAgos and their widespread associated proteins, suggesting an uncharted diversity of pAgo-derived immune systems.

Carbon quantum dots (CQDs) can be used to modify TiO

Type III CRISPR-Cas systems code for a multi-subunit ribonucleoprotein (RNP) complex that mediates DNA cleavage and synthesizes cyclic oligoadenylate (cOA) second messenger to confer anti-viral immunity. Both immune activities are to be activated upon binding to target RNA transcripts by their complementarity to crRNA, and autoimmunity avoidance is determined by extended complementarity between the 5ʹ-repeat tag of crRNA and 3ʹ-flanking sequences of target transcripts (anti-tag). However, as to how the strategy could achieve stringent autoimmunity avoidance remained elusive. In this study, we systematically investigated how the complementarity of the crRNA 5ʹ-tag and anti-tag (i.e., tag complementarity) could affect the interference activities (DNA cleavage activity and cOA synthesis activity) of Cmr-α, a type III-B system in Sulfolobus islandicus Rey15A. The results revealed an increasing suppression on both activities by increasing degrees of tag complementarity and a critical function of the 7(th) nucleotide of crRNA in avoiding autoimmunity. More importantly, mutagenesis of Cmr3α exerts either positive or negative effects on the cOA synthesis activity depending on the degrees of tag complementarity, suggesting that the subunit, coupling with the interaction between crRNA tag and anti-tag, function in facilitating immunity and avoiding autoimmunity in Type III-B systems.

Radio Frequency IDentification (RFID) technology is widely used in the inventory management of goods. Fast searching a subset consisting of some particular target tags is of practical importance for a variety of applications. Existing protocols are inefficient due to the low utilization of frame. In this paper, in order to improve the utilization of slot frame, we first generalize Cuckoo hash to extended Cuckoo hash which can use more than two hash functions. Then, we propose an extended CUckoo hash-based missing tag Detection (CUD) protocol, which can efficiently identify all missing tags within a predefined specific set. In CUD, the reader first uses the extended cuckoo hash to generate a fingerprint vector of the target tags. By using this vector, almost all target tags can be assigned a single slot and the majority of non-target tags can be filtered out. Furthermore, we optimize the parameters through theoretical analysis to minimize the execution time of the protocol. Extensive simulation results demonstrate that CUD is superior to the state-of-the-art protocols.

The durability of poly(vinyl chloride) (PVC) under long term exposure to heat and ultraviolet ray can be enhanced by adding an additive to improve its thermal stability. In this work, three types of cis-1,2-cyclohexanedicarboxylic acid di-mannitol ester (CAME)-based zinc metal alkoxides with different zinc contents (i.e., CAME-Zn-1, CAME-Zn-2, and CAME-Zn-3 containing 2:1, 1:1, and 1:2 CAME:Zn(O2CCH3)2 molar ratio, respectively) were characterized for the first time as PVC thermal stabilizer additives. Fourier-transform Infrared spectroscopy (FT-IR), elemental analyses results, and thermogravimetric analysis (TGA) results confirmed the formation of Zn-O bonds in CAME-Zn; that the Zn, C, H, and O contents on the as-prepared CAME-Zn are close to the targeted contents; and that CAME-Zn-2 has the highest decomposition temperature. Congo red tests, conductivity tests, and thermal aging tests subsequently revealed that relative to pure PVC, the thermal stability for the stabilized PVC was enhanced by adding thermal stabilizer in the order of ZnSt2/CaSt2 (1:1), CAME-Zn-3, CAME-Zn-1, and CAME-Zn-2. The best thermal stabilizer, i.e., CAME-Zn-2 can resist the color change from white to grey upon heated at 180 °C for up to 60 min. Torque rheological tests showed that CAME-Zn-2 contributes plasticizing effect to PVC. CAME-Zn-2 thus effectively contributes bi-functional roles, i.e., as thermal stabilizer and plasticizer to PVC.

Wave plates (WPs) are key components in optical polarization systems whose phase retardations and fast axis azimuths should be determined accurately. At present many measurement methods are based on light intensity detection which is inevitably affected by light intensity fluctuations so that the measurement accuracy is limited. What’s more, quite a few methods cannot measure WPs’ fast axis azimuths simultaneously. In this paper a method of simultaneously measuring the phase retardation and fast axis azimuth of arbitrary WP is proposed, which is based on equivalent component and phase detection. A rotatable half wave plate (HWP) and a retroreflector are used and its effect is equivalent to measuring an equivalent WP whose phase retardation is twice that of the measured wave plate, which is equivalent to doubling the measurement resolution. Phase detection is used to process the signals which means the measurement accuracy is better than that by usual light intensity detection. What’s more, the presented method eliminates the angle positioning errors of birefringent components in principle which exists in many present WP measurement methods. Finally, the measurement system setup is simple as well as the measurement process. The measurement formulae are deduced and corresponding WP measuring system is established. The error analysis shows that the system measurement uncertainty is about 3.9′ for the phase retardation and 5” for the fast axis azimuth. Experiment results and comparisons of quarter WPs and HWPs show that the presented method is in good agreement with other method. The phase retardation measurement repeatability is also good with a standard deviation about 2′.

Summary Type III CRISPR-cas systems initiate cyclic oligo-adenylate (cOA) signaling to initiate immune response. Previously, we identified that a membrane-associated DHH-DHHA1 family protein from Sulfolobus islandicus efficiently degrades cOA. Here, we provide detailed protocols for expression and purification of the protein from its native host and a cOA degradation assay with the purified enzyme. The methodology should be of interest for researchers studying Sulfolobus, membrane-associated proteins, or type III CRISPR-cas systems. For complete details on the use and execution of this protocol, please refer to Zhao et al. (2020)., Graphical Abstract, Highlights • Construct a Sulfolobus strain to express a membrane-associated DHH-DHHA1 protein (MAD) • Purify MAD by detergent treatment followed by chromatography • Analyze the degradation of type III CRISPR second messenger by MAD, Type III CRISPR-cas systems initiate cyclic oligo-adenylate (cOA) signaling to initiate immune response. Previously, we identified that a membrane-associated DHH-DHHA1 family protein from Sulfolobus islandicus efficiently degrades cOA. Here, we provide detailed protocols for expression and purification of the protein from its native host and a cOA degradation assay with the purified enzyme. The methodology should be of interest for researchers studying Sulfolobus, membrane-associated proteins, or type III CRISPR-cas systems.

Acquisition of spacers confers the CRISPR–Cas system with the memory to defend against invading mobile genetic elements. We previously reported that the CRISPR-associated factor Csa3a triggers CRISPR adaptation in Sulfolobus islandicus. However, a feedback regulation of CRISPR adaptation remains unclear. Here we show that another CRISPR-associated factor, Csa3b, binds a cyclic oligoadenylate (cOA) analog (5′-CAAAA-3′) and mutation at its CARF domain, which reduces the binding affinity. Csa3b also binds the promoter of adaptation cas genes, and the cOA analog enhances their binding probably by allosteric regulation. Deletion of the csa3b gene triggers spacer acquisition from both plasmid and viral DNAs, indicating that Csa3b acted as a repressor for CRISPR adaptation. Moreover, we also find that Csa3b activates the expression of subtype cmr-α and cmr-β genes according to transcriptome data and demonstrate that Csa3b binds the promoters of cmr genes. The deletion of the csa3b gene reduces Cmr-mediated RNA interference activity, indicating that Csa3b acts as a transcriptional activator for Cmr-mediated RNA interference. In summary, our findings reveal a novel pathway for the regulation of CRISPR adaptation and CRISPR–Cmr RNA interference in S. islandicus. Our results also suggest a feedback repression of CRIPSR adaptation by the Csa3b factor and the cOA signal produced by the Cmr complex at the CRISPR interference stage.

Sulfolobus islandicus codes for four DNA polymerases: three are of the B-family (Dpo1, Dpo2, and Dpo3), and one is of the Y-family (Dpo4). Western analysis revealed that among the four polymerases, only Dpo2 exhibited DNA damage-inducible expression. To investigate how these DNA polymerases could contribute to DNA damage tolerance in S. islandicus, we conducted genetic analysis of their encoding genes in this archaeon. Plasmid-borne gene expression revealed that Dpo2 increases cell survival upon DNA damage at the expense of mutagenesis. Gene deletion studies showed although dpo1 is essential, the remaining three genes are dispensable. Furthermore, although Dpo4 functions in housekeeping translesion DNA synthesis (TLS), Dpo2, a B-family DNA polymerase once predicted to be inactive, functions as a damage-inducible TLS enzyme solely responsible for targeted mutagenesis, facilitating GC to AT/TA conversions in the process. Together, our data indicate that Dpo2 is the main DNA polymerase responsible for DNA damage tolerance and is the primary source of targeted mutagenesis. Given that crenarchaea encoding a Dpo2 also have a low-GC composition genome, the Dpo2-dependent DNA repair pathway may be conserved in this archaeal lineage.

Japanese encephalitis virus (JEV) is a pathogen that causes severe vector-borne zoonotic diseases, thereby posing a serious threat to human health. Although JEV is potentially neurotropic, its pathogenesis and distribution in the host have not been fully elucidated. In this study, an infected mouse model was established using a highly virulent P3 strain of JEV. Immunohistochemistry and in situ hybridization, combined with anatomical imaging of the mouse brain, were used to dynamically localize the virus and construct three-dimensional (3D) images. Consequently, onset of mild clinical signs occurred in some mice at 3.5 d post JEV infection, while most mice displayed typical neurological signs at 6 d post-infection (dpi). Moreover, brain pathology revealed typical changes associated with non-suppurative encephalitis, which lasted up to 8 d. The earliest detection of viral antigen was achieved at 3 dpi in the thalamus and medulla oblongata. At 6 dpi, the positive viral antigen signals were mainly distributed in the cerebral cortex, olfactory area, basal ganglia, thalamus, and brainstem regions in mice. At 8 dpi, the antigen signals gradually decreased, and the localization of JEV tended to concentrate in the cerebrum and thalamus, while no viral antigen was detected in the brain at 21 dpi. In this model, the viral antigen was first expressed in the reticular thalamic nucleus (Rt), and the virus content is relatively stable. The expression of the viral antigen in the hippocampal CA2 region, the anterior olfactory nucleus, and the deep mesencephalic nucleus was high and persistent. The 3D images showed that viral signals were mostly concentrated in the parietal cortex, occipital lobe, and hippocampus, near the mid-sagittal plane. In the early stages of infection in mice, a large number of viral antigens were detected in denatured and necrotic neurons, suggesting that JEV directly causes neuronal damage. From the time of its entry, JEV is widely distributed in the central nervous system thereby causing extensive damage., Author summary There are many theories regarding the mechanism of entry of the Japanese encephalitis virus (JEV) into the nervous system. The inflammation cascade effect, resulting from the virus entering the central nervous system (CNS), is a major cause of brain injury in JEV patients. In this study, we found that the earliest point at which viral antigen was detected in the brain tissues following peripheral infection of JEV was at 3d. The virus was located in the nerve nuclei of the thalamus and medulla oblongata and, subsequently, viral antigens were found in the anterior olfactory nucleus. At 4 dpi, the virus was extensively distributed in the brain tissue, and at 6 d -8 d the viral antigen was widely distributed and highly concentrated. The viral concentration detected in the ventromedial thalamic nucleus (VM), deep mesencephalic nucleus (DpMe), and motor trigeminal nucleus (Mo5) remained high throughout the experiment. The hypertrophic nerve nuclei of JEV include the early anterior olfactory (AO) nucleus and the late hippocampal CA2 region. In the early stages of viral infection (6 dpi), the changes in viral antigen concentration and mortality rate were consistent. It was hypothesized that this stage represents the activation of viral proliferation and brain inflammation.

In this study, carbon quantum dots (CQDs) were synthesized by a low-cost and scalable approach and the oxygen functional groups were fine-tuned by chemical post-treatment. It was found that the CQDs could be applied as visible-light-responsive photocatalysts for activation of peroxymonosulfate (PMS) and remediation of aqueous organic dyes. Phenylhydrazine modified CQDs (CQDs-PH) presented high efficiency for degradation of methylene blue due to selective removal of carboxylic groups and inhibited recombination of photogenerated electron-hole pairs. The effects of catalyst dosage, species and concentrations of dyes, and initial pH values on the photodegradation efficiency were systematically investigated and the alkaline condition facilitates the separation of photoinduced charge carriers and promotes the dye decoloration. The reactive oxygen species produced in the photocatalysis were identified by radical quenching tests and the mechanism was elucidated. Superoxide radicals were generated from PMS activation via electron transfer from CQDs and played the primary role in organic oxidation. In addition, photogenerated holes on the valence band of CQDs also participated in the dye decomposition.

Type III CRISPR-Cas systems initiate an intracellular signaling pathway to confer immunity. The signalling pathway includes synthesis of cyclic oligo-adenylate (cOA) and activation of the RNase activity of type III accessory ribonuclease Csm6/Csx1 by cOA. After immune response, cOA should be cleared on time to avoid constant cellular RNA degradation. In this study, we show that the majority of cOA degradation activity is metal-dependent and associated to cell membrane in Sulfolobus islandicus. Further, a membrane-associated DHH-DHHA1 family nuclease (MAD) rapidly cleaves cOA and deactivates Csx1 ribonuclease. Compared to cellular ring nuclease, MAD uses a metal-dependent hydrolysis mechanism and is much more active. However, the subcellular organization may prevent it degrade nascent cOA. Together, the data suggest that MAD act as the second cOA degrader after ring nuclease to remove diffused redundant cOA.

Recently, Type III-A CRISPR-Cas systems were found to catalyze the synthesis of cyclic oligoadenylates (cOAs), a second messenger that specifically activates Csm6, a Cas accessory RNase and confers antiviral defense in bacteria. To test if III-B CRISPR-Cas systems could mediate a similar CRISPR signaling pathway, the Sulfolobus islandicus Cmr-α ribonucleoprotein complex (Cmr-α–RNP) was purified from the native host and tested for cOA synthesis. We found that the system showed a robust production of cyclic tetra-adenylate (c-A4), and that c-A4 functions as a second messenger to activate the III-B-associated RNase Csx1 by binding to its CRISPR-associated Rossmann Fold domain. Investigation of the kinetics of cOA synthesis revealed that Cmr-α–RNP displayed positively cooperative binding to the adenosine triphosphate (ATP) substrate. Furthermore, mutagenesis of conserved domains in Cmr2α confirmed that, while Palm 2 hosts the active site of cOA synthesis, Palm 1 domain serves as the primary site in the enzyme–substrate interaction. Together, our data suggest that the two Palm domains cooperatively interact with ATP molecules to achieve a robust cOA synthesis by the III-B CRISPR-Cas system.

Sulfolobus islandicus Rey15A encodes one Type I-A and two Type III-B systems, all of which are active in mediating nucleic acids interference. However, the effectiveness of each CRISPR system against virus infection was not tested in this archaeon. Here we constructed S. islandicus strains that constitutively express the antiviral immunity from either I-A, or III-B, or I-A plus III-B systems against SMV1 and tested the response of each host to SMV1 infection. We found that, although both CRISPR immunities showed a strong inhibition to viral DNA replication at an early stage of incubation, the host I-A CRISPR immunity gradually lost the control on virus proliferation, allowing accumulation of cellular viral DNA and release of a large number of viral particles. In contrast, the III-B CRISPR immunity showed a tight control on both viral DNA replication and virus particle formation. Furthermore, the SMV1 tolerance to the I-A CRISPR immunity did not result from the occurrence of escape mutations, suggesting the virus probably encodes an anti-CRISPR protein (Acr) to compromise the host I-A CRISPR immunity. Together, this suggests that the interplay between viral Acrs and CRISPR-Cas systems in thermophilic archaea could have shaped the stable virus-host relationship that is observed for many archaeal viruses.

The decline of proven conventional petroleum resources in the shallow portions of the Nanpu sag has shifted attention to the exploration of deep unconventional oil and gas resources there, especially the shale oil resources in the middle Eocene shales of the Shahejie Formation (MES shales). In this study, to assess the oil potential of the MES shales, we carried out a systematical geochemical and basin modeling characterization. The results show that the MES shales have low maturity and are at the early stage of hydrocarbon generation, with a mean vitrinite reflectance of 0.64% and low ratio of C27 18α(H)-22, 29, 30-trisnorneohopane to C27 17α(H)-22, 29, 30-trisnorhopane (0.44–1.18). The MES shales contain abundant organic matter (OM), with an average total organic carbon content of 2.21 wt %. Biomarker analyses show that the OM in the MES shales derived mainly from floating macrophyte and aquatic algae, microalgae and bacteria. The sapropelinite group in the MES shales is predominant in the organic macerals, and the kerogen is sapropelic and dominated by types I and II1. Biomarker analyses also reveal that the MES shales were deposited in the stratified water column as suggested by the presence of gammacerane, under relatively warm and wet climatic conditions. The anoxic conditions built up in the water column facilitated the OM preservation, although slight microbial degradation resulted in relatively high carbon preference index values. Modeling results based on geochemical data and basin analyses show that considerable quantities of hydrocarbons remain in the low-maturity MES shales. The preliminary estimate of the quantity of shale oil resources in the Gaoliu area of Nanpu sag is approximately 1.78 × 108 t. Nanpu sag have good shale oil potential, and shale oil exploration may be more feasible for MES shales, especially in the context of in-situ heating technology.

CRISPR–Cas (clustered regularly interspaced short palindromic repeats-CRISPR-associated) systems provide adaptive immunity against invasive nucleic acids guided by CRISPR RNAs (crRNAs) in archaea and bacteria. Type III CRISPR–Cas effector complexes show RNA cleavage and RNA-activated DNA cleavage activity, representing the only known system of dual nucleic acid interference. Here, we investigated the function of Cmr1 by genetic assays of DNA and RNA interference activity in the mutants and biochemical characterization of their mutated Cmr complexes. Three cmr1α mutants were constructed including ΔβΔ1α, Δβ1α-M1 and Δβ1α-M2 among which the last two mutants carried a double and a quadruple mutation in the first α-helix region of Cmr1α. Whereas the double mutation of Cmr1α (W58A and F59A) greatly influenced target RNA capture, the quadruple mutation almost abolished crRNA binding to Cmr1α. We found that Cmr2α-6α formed a stable core complex that is active in both RNA and DNA cleavage and that Cmr1α strongly enhances the basal activity of the core complex upon incorporation into the ribonucleoprotein complex. Therefore, Cmr1 functions as an integral activation module in III-B systems, and the unique occurrence of Cmr1 in III-B systems may reflect the adaptive evolution of type III CRISPR–Cas systems in thermophiles.

A new di-mannitol adipate ester-based zinc metal alkoxide (DMAE-Zn) was synthesized as a bi-functional poly(vinyl chloride) (PVC) thermal stabilizer for the first time. The materials were characterized with Fourier transform infrared spectroscopy (FT-IR) and thermogravimetric analysis (TGA). Characterization results confirmed the formation of Zn&ndash, O bonds in DMAE-Zn, and confirmed that DMAE-Zn had a high decomposition temperature and a low melting point. The thermal stability of DMAE-Zn on PVC also was tested by a conductivity test, a thermal aging test, and a UV-visible spectroscopy (UV-VIS) test. PVC stabilized by DMAE-Zn had a good initial color and excellent long-term stability. UV-VIS also showed that the conjugated structure in PVC stabilized by DMAE-Zn was almost all of the triene, suggesting that the addition of DMAE-Zn would suppress the formation of conjugated structures above tetraene. The dynamic processing performance of PVC samples tested by torque rheometer indicated that, having a good compatibility with PVC chains in the amorphous regions, DMAE-Zn contributed a good plasticizing effect to PVC. DMAE-Zn thus effectively demonstrates bi-functional roles, e.g., thermal stabilizers and plasticizers to PVC. Furthermore, FT-IR, a HCl absorption capacity test, and a complex ZnCl2 test were also used to verify the thermal stability mechanism of DMAE-Zn for PVC.

Thermal stabilizers, lubricant, and plasticizers are three crucial additives for processing poly(vinyl chloride) (PVC). In this study, a new mannitol stearate ester-based aluminum alkoxide (MSE-Al) was designed and synthesized as a novel additive for PVC. The thermal stability and processing performance of PVC stabilized by MSE-Al were evaluated by the Congo red test, conductivity measurement, thermal aging test, ultravioletevisible (UV&ndash, Vis) spectroscopy test, and torque rheometer test. Results showed that the addition of MSE-Al could not only markedly improve the long-term thermal stability of PVC, but also greatly accelerate the plasticizing and decrease the balance torque, which demonstrated that MSE-Al possessed a lubricating property. Thus, MSE-Al was demonstrated to be able to provide tri-functional additive roles, e.g., thermal stabilizer, plasticizer, and lubricant. The test results for the thermal stability of PVC indicated that the initial whiteness of PVC stabilized by MSE-Al was not good enough, thus the synergistic effect of MSE-Al with zinc stearates (ZnSt2) on the thermal stability of PVC was also investigated. The results showed that there is an appreciable synergistic effect between MSE-Al and ZnSt2. The thermal stabilization mechanism and synergism effect of MSE-Al with ZnSt2 are then discussed.

A new di-mannitol adipate ester-based zinc metal alkoxide (DMAE-Zn) was synthesized as a bi-functional PVC thermal stabilizer for the first time. The materials were characterized with Fourier-transform Infrared spectroscopy (FT-IR) and thermogravimetric analysis (TGA). Characterization results confirmed the formation of Zn-O bonds in DMAE-Zn; and that DMAE-Zn had a high decomposition temperature and a low melting point. The thermal stability of DMAE-Zn on PVC also was tested by conductivity test, thermal aging test, and UV-visible spectroscopy (UV-VIS) test. PVC stabilized by DMAE-Zn had a good initial color and excellent long-term stability. UV-VIS also showed that the conjugated structure in PVC stabilized by DMAE-Zn was almost all of the triene, suggesting that the addition of DMAE-Zn would suppress the formation of conjugated structures above tetraene. Dynamic processing performance of PVC samples tested by torque rheometer indicated that, having a good compatibility with PVC chains in the amorphous regions, DMAE-Zn contributed good plasticizing effect to PVC. DMAE-Zn thus effectively demonstrates bi-functional roles, e.g., thermal stabilizers and plasticizers to PVC. Furthermore, FT-IR, HCl absorption capacity test, and complex ZnCl2 test were also used to verify the thermal stability mechanism of DMAE-Zn for PVC.

Type III CRISPR-Cas systems code for a multi-subunit ribonucleoprotein (RNP) complex that mediates DNA cleavage and synthesizes cyclic oligoadenylate (cOA) second messenger to confer anti-viral immunity. Both immune activities are to be activated upon binding to target RNA transcripts by their complementarity to crRNA, and autoimmunity avoidance is determined by extended complementarity between the 5ʹ-repeat tag of crRNA and 3ʹ-flanking sequences of target transcripts (anti-tag). However, as to how the strategy could achieve stringent autoimmunity avoidance remained elusive. In this study, we systematically investigated how the complementarity of the crRNA 5ʹ-tag and anti-tag (i.e., tag complementarity) could affect the interference activities (DNA cleavage activity and cOA synthesis activity) of Cmr-α, a type III-B system in Sulfolobus islandicus Rey15A. The results revealed an increasing suppression on both activities by increasing degrees of tag complementarity and a critical function of the 7th nucleotide of crRNA in avoiding autoimmunity. More importantly, mutagenesis of Cmr3α exerts either positive or negative effects on the cOA synthesis activity depending on the degrees of tag complementarity, suggesting that the subunit, coupling with the interaction between crRNA tag and anti-tag, function in facilitating immunity and avoiding autoimmunity in Type III-B systems.

There is a wide demand for PVC products with microbiocidal property. For this purpose, additional antimicrobial agents have to be added. In this work, we synthesized pentaerythritol p-hydroxybenzoate ester-based zinc metal alkoxides (PHE-Zn) additive as highly efficient antibacterial agent, thermal stabilizer, plasticizer and lubricant for PVC. PHE-Zn was characterized by Fourier-transform infrared spectrometer (FT-IR), X-ray photoelectron spectroscopy (XPS), and elemental analysis. The results confirm the formation of R-O-Zn bonds and the molecular structure of PHE-Zn. The property enhancement of PVC by PHE-Zn were investigated by conductivity, oven aging, and torque rheological tests. PHE-Zn not only contributes to excellent initial whiteness and significant long-term thermal stability (without color change up to 90 min), but also improves plasticization and lubrication of PVC. In addition, using E. coli (E. coli) and Staphylococcus aureus (S. aureus) as typical microorganisms, the antibacterial properties of PHE-Zn stabilized PVC were tested. PVC with 4 phr PHE-Zn displayed high killing rates against E. coli and S. aureus (97.92% and 91.84%, respectively). Further experiments showed that PHE-Zn was a contact-active antibacterial agent. Moreover, the mechanisms for the enhanced thermal stability and biocidal property of PHE-Zn on PVC was explored.

Summary Type III CRISPR-Cas systems initiate an intracellular signaling pathway to confer immunity. The signaling pathway includes synthesis of cyclic oligo-adenylate (cOA) and activation of the RNase activity of type III accessory ribonuclease Csm6/Csx1 by cOA. After the immune response, cOA should be cleared on time to avoid constant cellular RNA degradation. In this study, we find a metal-dependent cOA degradation activity in Sulfolobus islandicus. The activity is associated with the cell membrane and able to accelerate cOA clearance at a high cOA level. Further, we show that a metal-dependent and membrane-associated DHH-DHHA1 family nuclease (MAD) rapidly cleaves cOA and deactivates Csx1 ribonuclease. The cOA degradation efficiency of MAD is much higher than the cellular ring nuclease. However, the subcellular organization may prevent it from degrading nascent cOA. Together, the data suggest that MAD acts as the second cOA degrader after the ring nuclease to remove diffused redundant cOA.