Structural Characterizations of the Dimeric Anti-HIV Antibody 2G12 and the HIV-2 Envelope Glycoprotein

More than thirty years after the discovery that Human Immunodeficiency Virus (HIV) was the causative agent of Acquired Immunodeficiency Syndrome (AIDS), the disease remains pandemic as long as no effective universal vaccine is found. Over 34 million individuals in the world are infected with the virus, and the vast majority of them have no access to the antiretroviral therapies that have largely reduced HIV to a chronic disease in the developed world. The first chapter of this thesis introduces the history of the virus. The key to the infectious mechanism of the virus lies in its envelope glycoprotein (Env), a trimeric spike on the viral surface that utilizes host T cell receptors for entry. Though HIV-1 Env is immunogenic, most infected patients do not mount an effective neutralizing antibody response against it. Broadly-neutralizing anti-Env antibodies (bNAbs) present in the serum of a minority of infected individuals are usually sufficient to prevent the progression to full blown AIDS. Thus, the molecular details of these bNAbs as well as the antibody-antigen interface are of prime interest for structural studies, as insight gained would contribute to the design of a more effective immunogen and potential vaccine candidate. The second chapter of this thesis describes the low-resolution crystal structure of one such antibody, 2G12 dimer, which targets a high mannose epitope on the surface of Env. Patients infected with HIV-2, a related virus with ~35% sequence identity in the Env region, can generally mount a robust antibody response sufficient for viral control for reasons still unknown. The final two chapters of this thesis focus on the first reported structural studies of HIV-2 Env, the molecular details of which may inform HIV-1 therapy and immunogen design

Diverse biological effects of glycosyltransferase genes from Tartary buckwheat

Background: Tartary buckwheat (Fagopyrum tataricum) is an edible cereal crop whose sprouts have been marketed and commercialized for their higher levels of anti-oxidants, including rutin and anthocyanin. UDP-glucose flavonoid glycosyltransferases (UFGTs) play an important role in the biosynthesis of flavonoids in plants. So far, few studies are available on UFGT genes that may play a role in tartary buckwheat flavonoids biosynthesis. Here, we report on the identification and functional characterization of seven UFGTs from tartary buckwheat that are potentially involved in flavonoid biosynthesis (and have varying effects on plant growth and development when overexpressed in Arabidopsis thaliana.) Results: Phylogenetic analysis indicated that the potential function of the seven FtUFGT proteins, FtUFGT6, FtUFGT7, FtUFGT8, FtUFGT9, FtUFGT15, FtUFGT40, and FtUFGT41, could be divided into three Arabidopsis thaliana functional subgroups that are involved in flavonoid biosynthesis of and anthocyanin accumulation. A significant positive correlation between FtUFGT8 and FtUFGT15 expression and anthocyanin accumulation capacity was observed in the tartary buckwheat seedlings after cold stress. Overexpression in Arabidopsis thaliana showed that FtUFGT8, FtUFGT15, and FtUFGT41 significantly increased the anthocyanin content in transgenic plants. Unexpectedly, overexpression of FtUFGT6, while not leading to enhanced anthocyanin accumulation, significantly enhanced the growth yield of transgenic plants. When wild-type plants have only cotyledons, most of the transgenic plants of FtUFGT6 had grown true leaves. Moreover, the growth speed of the oxFtUFGT6 transgenic plant root was also significantly faster than that of the wild type. At later growth, FtUFGT6 transgenic plants showed larger leaves, earlier twitching times and more tillers than wild type, whereas FtUFGT15 showed opposite results. Conclusions: Seven FtUFGTs were isolated from tartary buckwheat. FtUFGT8, FtUFGT15, and FtUFGT41 can significantly increase the accumulation of total anthocyanins in transgenic plants. Furthermore, overexpression of FtUFGT6 increased the overall yield of Arabidopsis transgenic plants at all growth stages. However, FtUFGT15 shows the opposite trend at later growth stage and delays the growth speed of plants. These results suggested that the biological function of FtUFGT genes in tartary buckwheat is diverse

Contrastive Modules with Temporal Attention for Multi-Task Reinforcement Learning

In the field of multi-task reinforcement learning, the modular principle, which involves specializing functionalities into different modules and combining them appropriately, has been widely adopted as a promising approach to prevent the negative transfer problem that performance degradation due to conflicts between tasks. However, most of the existing multi-task RL methods only combine shared modules at the task level, ignoring that there may be conflicts within the task. In addition, these methods do not take into account that without constraints, some modules may learn similar functions, resulting in restricting the model's expressiveness and generalization capability of modular methods. In this paper, we propose the Contrastive Modules with Temporal Attention(CMTA) method to address these limitations. CMTA constrains the modules to be different from each other by contrastive learning and combining shared modules at a finer granularity than the task level with temporal attention, alleviating the negative transfer within the task and improving the generalization ability and the performance for multi-task RL. We conducted the experiment on Meta-World, a multi-task RL benchmark containing various robotics manipulation tasks. Experimental results show that CMTA outperforms learning each task individually for the first time and achieves substantial performance improvements over the baselines.Comment: This paper has been accepted at NeurIPS 2023 as a poste

A replicative recombinant HPV16 E7 expression virus upregulates CD36 in C33A cells

ObjectiveIn past decades, the role of high-risk HPV (HR-HPV) infection in cancer pathogenesis has been extensively studied. The viral E7 protein expressed in pre-malignant cells has been identified as an ideal target for immunological intervention. However, the cultivation of HPV in vitro remains a significant challenge, as well as the lack of methods for expressing the HPV E7 protein and generating replication-competent recombinant viral particles, which posed a major obstacle to further exploration of the function and carcinogenic mechanisms of the E7 oncoprotein. Therefore, it is imperative to investigate novel methodologies to construct replication-competent recombinant viral particles that express the HPV E7 protein to facilitate the study of its function.MethodsWe initiated the construction of recombinant viral particles by utilizing the ccdB-Kan forward/reverse screening system in conjunction with the Red/ExoCET recombinant system. We followed the infection of C33A cells with the obtained recombinant virus to enable the continuous expression of HPV16 E7. Afterwards, the total RNA was extracted and performed transcriptome sequencing using RNA-Seq technology to identify differentially expressed genes associated with HPV-induced oncogenicity.ResultsWe successfully established replicative recombinant viral particles expressing HPV16 E7 stably and continuously. The C33A cells were infected with recombinant viral particles to achieve overexpression of the E7 protein. Subsequently, RNA-Seq analysis was conducted to assess the changes in host cell gene expression. The results revealed an upregulation of the CD36 gene, which is associated with the HPV-induced oncogenic pathways, including PI3K-Akt and p53 signaling pathway. qRT-PCR analysis further identified that the upregulation of the CD36 gene due to the expression of HPV16 E7.ConclusionThe successful expression of HPV16 E7 in cells demonstrates that the replicated recombinant virus retains the replication and infection abilities of Ad4, while also upregulating the CD36 gene involved in the PI3K-Akt signaling and p53 pathways, thereby promoting cell proliferation. The outcome of this study provides a novel perspective and serves as a solid foundation for further exploration of HPV-related carcinogenesis and the development of replicative HPV recombinant vaccines capable of inducing protective immunity against HPV

Approach to Higher Wheat Yield in the Huang-Huai Plain: Improving Post-anthesis Productivity to Increase Harvest Index

Both increased harvest index (HI) and increased dry matter (DM) are beneficial to yield; however, little is known about the priority of each under different yield levels. This paper aims to determine whether HI or DM is more important and identify the physiological attributes that act as indicators of increased yield. Two field experiments involving different cultivation patterns and water-nitrogen modes, respectively, were carried out from 2013 to 2016 in Huang-Huai Plain, China. Plant DM, leaf area index (LAI), and radiation interception (RI) were measured. Increased yield under low yield levels <7500 kg ha-1 was attributed to an increase in both total DM and HI, while increases under higher yield levels >7500 kg ha-1 were largely dependent on an increase in HI. Under high yield levels, HI showed a significant negative correlation with total DM and a parabolic relationship with net accumulation of DM during filling. Higher net accumulation of DM during filling helped slow down the decrease in HI, thereby maintaining a high value. Moreover, net DM accumulation during filling was positively correlated with yield, while post-anthesis accumulation showed a significant linear relationship with leaf area potential (LAP, R2 = 0.404–0.526) and radiation interception potential (RIP, R2 = 0.452–0.576) during grain filling. These findings suggest that the increase in LAP and RIP caused an increase in net DM accumulation after anthesis. Under DM levels >13,000 kg ha-1 at anthesis, maintaining higher LAI and RI in lower layers during grain formation contributed to higher yield. Furthermore, the ratio of upper- to lower-layer RI showed a second-order curve with yield during filling, with an increase in the optimal range with grain development. Pre-anthesis translocation amount, translocation ratios and contribution ratios also showed second-order curves under high yield levels, with optimal values of 3000–4500 kg ha-1, 25–35, and 30–50%, respectively. These results confirm the importance of HI in improving the yield, thereby providing a theoretical basis for wheat production in the Huang-Huai Plain

Structural Basis for Enhanced HIV-1 Neutralization by a Dimeric Immunoglobulin G Form of the Glycan-Recognizing Antibody 2G12

The human immunoglobulin G (IgG) 2G12 recognizes high-mannose carbohydrates on the HIV type 1 (HIV-1) envelope glycoprotein gp120. Its two antigen-binding fragments (Fabs) are intramolecularly domain exchanged, resulting in a rigid (Fab)_2 unit including a third antigen-binding interface not found in antibodies with flexible Fab arms. We determined crystal structures of dimeric 2G12 IgG created by intermolecular domain exchange, which exhibits increased breadth and >50-fold increased neutralization potency compared with monomeric 2G12. The four Fab and two fragment crystalline (Fc) regions of dimeric 2G12 were localized at low resolution in two independent structures, revealing IgG dimers with two (Fab)_2 arms analogous to the Fabs of conventional monomeric IgGs. Structures revealed three conformationally distinct dimers, demonstrating flexibility of the (Fab)_2-Fc connections that was confirmed by electron microscopy, small-angle X-ray scattering, and binding studies. We conclude that intermolecular domain exchange, flexibility, and bivalent binding to allow avidity effects are responsible for the increased potency and breadth of dimeric 2G12

Studies in Chinese lexicology: investigations into the Xiang dialect

© 1992 Dr. Yunji WuAny speaker of a non-standard Chinese dialect has some familiarity with a number of written and spoken varieties of the Chinese language, both dialectal and standard. The knowledge of these varieties, and of relationships between them, is an essential characteristic of a dialect speaker's linguistic competence in Chinese, and the complexity of this knowledge presents one of the greatest difficulties for the linguist in charting the synchronic structures and diachronic development of Chinese. The challenge begins for the native speaker, who must negotiate their way through this linguistic milieu. It extends to and dominates the work of contemporary Chinese dialectologists, the products of whose research cannot be properly understood unless one has a clear grasp of both the Chinese dialect speaker's complex linguistic world, and also the traditional means by which Chinese researchers have developed to represent this world. This thesis attempts to tackle this challenge through a discussion of problems concerning the status and organization of the Changsha dialect. The main focus is upon the formative: the monosyllabic morpheme, which, from the perspective of the Chinese speaker, is the fundamental unit in word formation. The arrangement and rearrangement of phonological-semantic relationships between formatives within and across dialects will be explored through a discussion of types of local words (Chapter 3), interactions between different varieties, both written and spoken" standard and local (Chapter 4), and the problem of representation of local words in characters (Chapter 5). Chapters 1 and 2 provide a general introduction to these issues, and also to the linguistic situation of Hunan, the province of which Changsha is the capital

Structural Basis for Enhanced Hiv-1 Neutralization by a Dimeric Immunoglobulin G Form of the Glycan-Recognizing Antibody 2G12

The human immunoglobulin G (IgG) 2G12 recognizes high-mannose carbohydrates on the HIV type 1 (HIV-1) envelope glycoprotein gp120. Its two antigen-binding fragments (Fabs) are intramolecularly domain exchanged, resulting in a rigid (Fab)2 unit including a third antigen-binding interface not found in antibodies with flexible Fab arms. We determined crystal structures of dimeric 2G12 IgG created by intermolecular domain exchange, which exhibits increased breadth and >50-fold increased neutralization potency compared with monomeric 2G12. The four Fab and two fragment crystalline (Fc) regions of dimeric 2G12 were localized at low resolution in two independent structures, revealing IgG dimers with two (Fab)2 arms analogous to the Fabs of conventional monomeric IgGs. Structures revealed three conformationally distinct dimers, demonstrating flexibility of the (Fab)2-Fc connections that was confirmed by electron microscopy, small-angle X-ray scattering, and SPR binding studies. We conclude that intermolecular domain exchange, flexibility, and bivalent binding to allow avidity effects are responsible for the increased potency and breadth of dimeric 2G12. In addition, we present this as the first known crystal structure of an IgG dimer

Simultaneous Event-Triggered Fault Detection and Estimation for Stochastic Systems Subject to Deception Attacks

In this paper, a synthesized design of fault-detection filter and fault estimator is considered for a class of discrete-time stochastic systems in the framework of event-triggered transmission scheme subject to unknown disturbances and deception attacks. A random variable obeying the Bernoulli distribution is employed to characterize the phenomena of the randomly occurring deception attacks. To achieve a fault-detection residual is only sensitive to faults while robust to disturbances, a coordinate transformation approach is exploited. This approach can transform the considered system into two subsystems and the unknown disturbances are removed from one of the subsystems. The gain of fault-detection filter is derived by minimizing an upper bound of filter error covariance. Meanwhile, system faults can be reconstructed by the remote fault estimator. An recursive approach is developed to obtain fault estimator gains as well as guarantee the fault estimator performance. Furthermore, the corresponding event-triggered sensor data transmission scheme is also presented for improving working-life of the wireless sensor node when measurement information are aperiodically transmitted. Finally, a scaled version of an industrial system consisting of local PC, remote estimator and wireless sensor node is used to experimentally evaluate the proposed theoretical results. In particular, a novel fault-alarming strategy is proposed so that the real-time capacity of fault-detection is guaranteed when the event condition is triggered