Successive Generations Research Articles

Cloning genes of p49, p72, pe199l, pe248r and cd2vinto <i>Pichia Pastoris</i> gs115

African swine fever (ASF), caused by the African swine fever virus (ASFV), poses a significant threat to pig populations worldwide. ASFV is a double-stranded DNA virus. In recent years, 54 structural proteins and more than 100 proteins have been involved in viral infection found in macrophages of diseased pigs. Among these, p49 and p72 are essential capsid proteins crucial for forming the viral capsid. Additionally, pE199L and pE248R, located in the inner viral membrane, are critical for membrane fusion, a necessary step for viral entry into host cells. Another important player is CD2v, a type I transmembrane protein involved in the infection process. In this research, to develop subunit vaccines against ASFV, we focused on cloning the genes encoding these five proteins—p49, p72, pE199L, pE248R, and CD2v—into the pPIC9K plasmid for expression in the Pichia pastoris GS115 yeast strain. The viral genomic DNA was extracted from blood samples of infected pigs, and the genes encoding the five proteins were successfully amplified using Phusion PCR. The PCR products of each gene were then digested with EcoRI and NotI restriction enzymes and ligated into the pPIC9K plasmid. After that, we transformed the recombinant plasmids into Escherichia coli DH5α for amplification and purification. The plasmids were subsequently linearized with SalI and introduced into P. pastoris GS115 through electroporation. The selection of appropriate media and PCR analysis of the genomic DNA confirmed the successful generation of five recombinant P. pastoris GS115 strains. This work paves the way for the development of a recombinant protein vaccine against ASF by using the Pichia pastoris GS115 in the future.

Deep Learning-Assisted Design of de novo Protein Binders Targeting Hepatitis C Virus E2 Protein

Hepatitis C virus is a grievous disease with an increased mortality rate worldwide. Chemical-based medications possess deleterious side effects and are considered inefficient in combating viral infections. Advanced therapeutic strategies are being examined with increased specificity against viral proteins such as designing highly regular proteins facilitating the development of highly effective inhibitors. Here, we present for the first time, the use of deep learning-based large language protein model ProtGPT2 with a unique strategy to design novel therapeutic binders that have the potential to mimic host receptors and inhibit the viral protein, especially the HCV envelope glycoprotein E2 for clinically relevant genotype 1a and 1b. We generated five de novo proteins for each host receptor that mimic the human receptors, based on the interacting residues which were identified by the tools of the PDBSum database in the docked host-E2 complexes generated with the ClusPro web server. The Root Mean Square Deviation score revealed that each de novo designed binder exhibited high similarity with the human receptors indicating a successful generation. Furthermore, multiple interactions were observed between these de novo designed proteins and E2 protein, emphasizing the potential of these de novo designed proteins as significant inhibitors. A comparative analysis of molecular docking between human interacting partners and de novo designed proteins revealed that de novo proteins, such as CD81-D1 and CLDN-D4, are the most effective inhibitors having the lowest binding energy when interacting with the most conserved regions of the E2 protein. These generated proteins may inhibit the interaction of E2 with CD81 and CLDN host receptors.

Démence Précoce: Historical Conceptualization of the Concept

Schizophrenia, like many mental disorders, was historically viewed as madness or demonic possession until the 18th century when psychiatry began to emerge as a scientific discipline. French psychiatrist Bénédict Morel played a pivotal role in this transformation by coining the term "<i>démence précoce</i>." He used this term to describe a set of clinical features observed in schizophrenia, suggesting that it was due to an inherent biological defect that progressively worsened through successive generations. German psychiatrist Emil Kraepelin further refined Morel's ideas by developing the concept of "<i>dementia praecox</i>". He described a cluster of symptoms and signs characterized by a specific course and outcome, emphasizing the chronic and deteriorative nature of the illness. Kraepelin's work laid the groundwork for modern understanding, emphasizing the classification of psychiatric conditions based on symptom clusters, progression, and outcomes. In the early 20th century, Swiss psychiatrist Eugen Bleuler introduced the term "<i>schizophrenia</i>", marking a significant shift in the conceptualization of the disorder. He viewed it not as a single disease but as a group of related disorders, which he called the "group of schizophrenias." Bleuler focused on the splitting of cognitive functions—such as thinking, feeling, and behavior—recognizing a broader spectrum of symptoms beyond the purely degenerative model proposed by Kraepelin. The evolution of schizophrenia's conceptualization reflects broader developments in neuropsychiatry, neuropsychopharmacology, and neuroscience. Advances in these fields have refined diagnostic criteria, understanding of neurobiological underpinnings, and treatment approaches. However, the journey to fully understanding schizophrenia is ongoing. The complexities of its causes, manifestations, and treatments mean that its history—and the final chapter of its understanding—remains to be written, as new research continues to challenge and expand upon existing knowledge.

Room-temperature waveguide integrated quantum register in a semiconductor photonic platform

Quantum photonic integrated circuits are reshaping quantum networks and sensing by providing compact, efficient platforms for practical quantum applications. Despite continuous breakthroughs, integrating entangled registers into photonic devices on a CMOS-compatible platform presents significant challenges. Herein, we present single electron-nuclear spin entanglement and its integration into a silicon-carbide-on-insulator (SiCOI) waveguide. We demonstrate the successful generation of single divacancy electron spins and near-unity spin initialization of single 13C nuclear spins. Both single nuclear and electron spin can be coherently controlled and a maximally entangled state with a fidelity of 0.89 has been prepared under ambient conditions. Based on the nanoscale positioning techniques, the entangled quantum register has been further integrated into SiC photonic waveguides for the first time. We find that the intrinsic optical and spin characteristics of the register are well preserved and the fidelity of the entangled state remains as high as 0.88. Our findings highlight the promising prospects of the SiCOI platform as a compelling candidate for future scalable quantum photonic applications.

Experimental Evolution Induced by Maternal Post-copulatory Factors in Drosophila.

Experimental evolution is a powerful approach to study the mechanisms underlying the adaptation of selected characters under the conditions chosen in the laboratory. Drosophila melanogaster is a species frequently used to investigate the experimental evolution of characters, especially those related to reproduction. Recent intra-generational studies showed that cis-vaccenyl acetate (cVa), a sex pheromone transferred with bacteria on eggs by females either 1day (D1) or 5days (D5) after copulation, differentially affected the behavior and pheromone release in adult males emerging from these eggs. Here, we extended this finding to determine whether this alternative egg exposure repeated over many generations could affect a larger set of reproduction-related characters in both sexes. To test the repetitive effects of maternal D1 or D5 post-copulatory factors, we carried out an experimental selection procedure consisting of exposing eggs during 40 successive generations to D1 or D5 maternal post-copulatory factors. We compared cVa and cuticular pheromones, courtship and mating behaviors, and fecundity at different generations in flies of D1 and D5 lines. Based on findings obtained at earlier generations, we also determined survival, bacterial composition and gene expression in adults. Some of these complex traits significantly diverged between D1 and D5 lines indicating that maternal post-copulatory factors transmitted to eggs can influence adult life history traits.

Generation of viable hypomorphic and null mutant plants via CRISPR-Cas9 targeting mRNA splicing sites.

Genetic analysis is important for modern plant molecular biology, and in this regard, the existence of specific mutants is crucial. While genome editing technologies, particularly CRISPR-Cas9, have revolutionized plant molecular biology by enabling precise gene disruption, knockout methods are ineffective for lethal genes, necessitating alternatives like gene knockdown. This study demonstrates the practical generation of a hypomorphic mutant allele, alongside severe null mutant alleles, via the targeting of mRNA splicing sites using CRISPR-Cas9. The Arabidopsis HIGH PLOIDY 2 (HPY2) encodes a yeast NSE2 ortholog, part of the conserved eukaryotic SMC5/6 complex, with SUMO E3 ligase activity essential for cell cycle progression and plant development. Loss-of-function HPY2 mutants exhibit severe dwarfism and seedling lethality, making functional analysis challenging. To overcome these limitations, we created HPY2 knockdown mutants as novel tools to investigate gene function. Of the three mutant alleles, the hpy2-cr1 and hpy2-cr2 mutants resembled the existing severe hpy2-1 allele, both harboring a single base pair insertion in one exon, causing significant root shortening and seedling lethality. In contrast, the hypomorphic mutant hpy2-cr3, which has a five bp deletion at an intron-exon junction, showed relatively longer root growth and survived until the reproductive stage. RT-PCR analysis of hpy2-cr3 revealed atypical mRNAs producing truncated polypeptides that retained some HPY2 function, explaining the milder phenotype. These results establish the successful generation of novel hypomorphic mutant alleles critical for studying the lethal gene HPY2, and demonstrate the usefulness of CRISPR-Cas9 for producing viable hypomorphic mutants for investigating complex genetic interactions.

Wild again: recovery of a beneficial Cannabis seed endophyte from low domestication genotypes

BackgroundBeyond carrying the plant embryo, seeds harbour intricate microbial communities whose transmission across successive plant generations can significantly influence the ecological and evolutionary dynamics of plant–microbe symbioses. The process of plant domestication has potential repercussions in genes involved in plant-microbiome interactions. However, the extent to which breeding can impact the seed microbiome is sparsely explored. Cannabis is a high-value crop but sparsely subjected to agricultural innovations established in other crop species during the last century. Here, we conduct a large-scale analysis of the bacterial seed microbiome of Cannabis across different domestication grades and investigate the potential of seed-associated endophytes as plant growth-promoting agents under both controlled and field conditions.ResultsAnalysis of Cannabis seed endophyte composition and diversity across 46 plant genotypes revealed 813 different bacterial genera with a predominance of Gammaproteobacteria, Bacilli, Actinobacteria and Alphaproteobacteria but a genotype-specific microbiome. The assessment of domestication and breeding on microbial assembly revealed a higher bacterial diversity in low domestication genotypes (Shannon index, H′: 1.21 vs. 1.05) and a higher homogeneity in bacterial composition caused by line development. Further, a seed bacterial isolate (Bacillus frigoritolerans C1141) associated with low domestication genotypes, and with genes associated with bio-fertilization, bioremediation and phytohormone production, increased plant growth by 42.3% at the time of harvest, under field conditions.ConclusionThis study addresses critical knowledge gaps related to the assembly of the Cannabis seed-endophytic microbiome. It reveals that Cannabis breeding is linked to alterations of seed microbial communities, which potentially led to the loss of bacteria with functional significance. These results highlight the importance of preserving seed microbiomes in plant breeding to support sustainable plant health and growth enhancement in Cannabis.CAHvc7HPtw2foEoMit4yw-Video

Strategic insights into the cultivation of pancreatic cancer organoids from endoscopic ultrasonography-guided biopsy tissue

BACKGROUND The frequent suboptimal efficacy of endoscopic ultrasound-guided fine-needle biopsy (EUS-FNB) to culture pancreatic cancer (PC) organoids (PCOs) poses a major challenge in the advancement of personalized medicine for advanced PC. AIM To explore how to obtain appropriate puncture tissues from EUS-FNB and optimize the strategy for efficiently constructing PCOs, providing an efficient tool for the advancement of personalized medicine. METHODS Patients who underwent EUS-FNB for the diagnosis of PC tissue were prospectively enrolled. We refined the endoscopic biopsy procedures and organoid cultivation techniques. All tissue specimens verified by on-site pathological assessment were cultured in a semi-suspended medium in a microfluidic environment. We assessed differences in PCOs cultured beyond and below five generations examining patient demographics, specimen and organoid attributes, and the sensitivity of organoids to a panel of clinical drugs through cell viability assays. RESULTS In this study, 16 patients with PC were recruited, one sample was excluded because onsite cytopathology showed no tumor cells. Successful organoid generation occurred in 93.3% (14 of 15) of the EUS-FNB specimens, with 60% (9 of 15) sustaining over five generations. Among these patients, those with a history of diabetes, familial cancer, or larger tumors exhibited enhanced PCO expandability. The key factors influencing long-term PCOs expansion included initial needle sample quality (P = 0.005), rapid initiation of organoid culture post-isolation (P ≤ 0.001), and high organoid activity (P = 0.031). Drug sensitivity analysis revealed a partial response in two patients following therapeutic intervention and surgery and stable disease in four patients, indicating a moderate correlation between organoid response and clinical outcomes. CONCLUSION Optimal initial needle sampling, rapid and precise biopsy sample processing, process isolated samples as soon as possible, and sufficient cellular material are crucial for successful cultivating PCOs. High organoid activity is an important factor in maintaining their long-term expansion, which is essential for shortening the time of drug sensitivity analysis and is the basis of PC research.

Targeted engineering of camelina and pennycress seeds for ultrahigh accumulation of acetyl-TAG

Acetyl-TAG (3-acetyl-1,2-diacylglycerol), unique triacylglycerols (TAG) possessing an acetate group at the sn-3 position, exhibit valuable properties, such as reduced viscosity and freezing points. Previous attempts to engineer acetyl-TAG production in oilseed crops did not achieve the high levels found in naturally producing Euonymus seeds. Here, we demonstrate the successful generation of camelina and pennycress transgenic lines accumulating nearly pure acetyl-TAG at 93 mol% and 98 mol%, respectively. These ultrahigh acetyl-TAG synthesizing lines were created using gene-edited FATTY ACID ELONGASE1 (FAE1) mutant lines as an improved genetic background to increase levels of acetyl-CoA available for acetyl-TAG synthesis mediated by the expression of EfDAcT, a high-activity diacylglycerol acetyltransferase isolated from Euonymus fortunei. Combining EfDAcT expression with suppression of the competing TAG-synthesizing enzyme DGAT1 further enhanced acetyl-TAG accumulation. These ultrahigh levels of acetyl-TAG exceed those in earlier engineered oilseeds and are equivalent or greater than those in Euonymus seeds. Imaging of lipid localization in transgenic seeds revealed that the low amounts of residual TAG were mostly confined to the embryonic axis. Similar spatial distributions of specific TAG and acetyl-TAG molecular species, as well as their probable diacylglycerol (DAG) precursors, provide additional evidence that acetyl-TAG and TAG are both synthesized from the same tissue-specific DAG pools. Remarkably, this ultrahigh production of acetyl-TAG in transgenic seeds exhibited minimal negative effects on seed properties, highlighting the potential for production of designer oils required for economical biofuel industries.

Deep learning-driven super-resolution in Raman hyperspectral imaging: Efficient high-resolution reconstruction from low-resolution data

Deep learning (DL) has become an indispensable tool in hyperspectral data analysis, automatically extracting valuable features from complex, high-dimensional datasets. Super-resolution reconstruction, an essential aspect of hyperspectral data, involves enhancing spatial resolution, particularly relevant to low-resolution hyperspectral data. Yet, the pursuit of super-resolution in hyperspectral analysis is fraught with challenges, including acquiring ground truth high-resolution data for training, generalization, and scalability. The pressing issue of extended spectral acquisition times, notably for high-resolution scans, is a significant roadblock in hyperspectral imaging. Super-resolution methods offer a promising solution by providing higher spatial resolution data to expedite data collection and yield more efficient outcomes. This paper delves into a practical application of these concepts using Raman imaging, where spectral acquisition times can be prohibitively long. In this context, DL-based super-resolution models demonstrate their efficacy by predicting and reconstructing high-resolution Raman data from low-resolution input, eliminating the need for resource-intensive high-resolution scans. While previous work often relied on substantial high-resolution datasets, this study showcases the ability to achieve similar outcomes even with limited data, presenting a more practical and cost-effective approach. The results offer a glimpse into the transformative potential of this technology to streamline hyperspectral imaging applications by saving valuable time and resources through the successful generation of high-resolution data from low-resolution inputs.

HOST COUNTRY LANGUAGE ACQUISITION AMONG FIRST-GENERATION YAGLIDERE IMMIGRANTS IN THE USA AND THE MAINTENANCE OF THE TURKISH LANGUAGE IN SUCCESSIVE GENERATIONS VIA SOCIAL NETWORKS

This article investigated the effect of the social networks of Yaglidere Turkish immigrants on language acquisition and the maintenance of heritage languages by successive generations in the USA. The study was conducted in the cities of Patterson and Burlington between January 2018 and July 2018. This research used qualitative research methods to analyze ethnographic interviews and participant observations. Data were collected using participant observation and informal talks with immigrants conducted in different informal settings such as participants’ houses, their restaurants, and other cafes in Burlington and Paterson. The interviews lasted from 40 to 90 minutes. Questions were open-ended and focused on the role of social networks in the acquisition of English language skills. In order to analyze the resulting data, the audio recordings were transcribed and translated into English. The interviews were first transcribed into Microsoft Word files and then imported into Maxqda, a software program for qualitative analysis, where the data were coded for study. The results indicated that even though social networks have destructive effects among first-generation immigrants, they have undeniable constructive effects on language maintenance in other generations. So, what is destructive for the first generation turns into constructive for the other generations.

Emergence of dauer larvae in Caenorhabditis elegans disrupts continuity of host-microbiome interactions.

Nematodes are common in most terrestrial environments, where populations are often known to undergo cycles of boom and bust. Useful in such scenarios, nematodes present developmental programs of diapause, giving rise to stress-resistant larvae and enabling dispersal in search of new resources. Best studied in Caenorhabditis elegans, stress resistant dauer larvae emerge under adverse conditions, primarily starvation, and migrate to new niches where they can resume development and reproduce. C. elegans is a bacterivore but has been shown to harbor a persistent and characteristic gut microbiome. While much is known about the gut microbiome of reproducing C. elegans, what dauers harbor is yet unknown. This is of interest, as dauers are those that would enable transmission of microbes between nematode generations and geographical sites, maintaining continuity of host-microbe interactions. Using culture-dependent as well as sequencing-based approaches we examined the gut microbiomes of dauers emerging following population growth on ten different natural-like microbially diverse environments as well as on two defined communities of known gut commensals and found that dauers were largely devoid of gut bacteria. These results suggest that host gut-microbiome interactions in C. elegans are not continuous across successive generations and may reduce the likelihood of long-term worm-microbe coevolution.

Virus-Induced Gene Silencing Simultaneously Exploits ‘Attract and Kill’ Traits in Plants and Insects to Manage Huanglongbing

Abstract The vector-borne disease Huanglongbing (HLB) causes severe economic losses to citrus production worldwide with no available cure. Herein, we applied virus-induced gene silencing technology to engineer citrus that preferentially attracted and specifically killed Diaphorina citri, the vector associated with HLB. We engineered the infectious citrus tristeza virus (CTV-T36) clone to carry three truncated genes. The triple construct (CTV-tAwd-tWnt-tPDS) produces small interfering RNAs (siRNAs) against phytoene desaturase, PDS, to yield a phenotype with visual, olfactory, and gustatory cues that preferentially attracted D. citri. In addition, siRNAs targeted two genes related to flight in D. citri, abnormal wing disc (DcAwd) and wingless (DcWnt), that caused wing malformations and decreased survival in psyllids that fed on plants inoculated with the engineered virus. During two successive generations, D. citri reared on CTV-tAwd-tWnt-tPDS-inoculated plants exhibited higher mortality across life stages as well as reduced fecundity and fertility as compared with those reared on non-infected plants or CTV-wt-inoculated plants. Furthermore, CTV-tAwd-tWnt-tPDS-inoculated plants shortened the lifespan of D. citri by more than 20 days. Morphological abnormalities were noted in those adults that did successfully emerge on plants inoculated with CTV-tAwd-tWnt-tPDS, including cocked wings with a bowl-shaped depression and/or a convex shape. Phloem sap from CTV-tAwd-tWnt-tPDS-inoculated plants decreased the survival of D. citri adults, confirming that siRNAs were present in the sap of these plants. Collectively, we provide proof of concept for a novel variant of the attract-and-kill method where the cultivated crop is potentially transformed into a hyper-attractive population and transmission sink for a phytopathogen vector.

Nano-priming of Vigna radiata seeds with opuntia stricta-derived γFe2O3 nanoparticles

The experimental investigation aimed to enhance sustainable and biocompatible nano-agriculture by examining the possible effects of biosynthesized nanoparticles (γFe2O3 NPs) on seed growth and growth parameters of Vigna radiata. The outcomes of various characterization procedures such as FTIR, XRD, UV, and Fe-SEM were used to determine the successful generation of γFe2O3 NPs (nanoparticles), which were capped with multiple phytoconstituents contained in the Opuntia stricta extract. γFe2O3 NPs were synthesised, as evidenced by the peak's presence, having a mean crystallite size of 13.9 nm, and were successfully detected using X-ray diffraction (XRD). Crucial Energy Dispersive X-ray Spectroscopy (EDS) testing also yielded unambiguous proof of the element iron's presence. Raman spectroscopy was used to confirm the synthesis of pure maghemite nanoparticles. Wide areas around 350, 500, 670, and 1330 cm-1 values verify the creation of pure maghemite. TGA analysis was performed to check the stability of nanoparticles in which the residual weight was found to be 87.62%. TEM images confirm the spherical shape, and the mean size of the nanoparticles was found to be 12.28 nm. Through a number of tests, the analysis showed that the nanoparticles have a strong antioxidant capacity. This experiment aimed to determine how biogenic γFe2O3 Nanoparticles affected Vigna radiata seed germination. Measurements of shoot and root lengths, seed vigour index, germination rates, mean daily germination, and peak value (PV) were all included of the assessment. There was a total of four distinct γFe2O3 NPs concentrations examined (25, 50, 75, and 100 μg/mL). The penetration of the γFe2O3 NPs affects the germination and growth of the Vigna radiata seeds. When compared to untreated seeds, the germination index values for Vigna radiata seeds that underwent effective treatment with γFe2O3 NPs significantly improved.

Belief Formation: A Cultural Evolutionary Lens

This paper investigates changes in belief and their evolution in relation to the paradigm of cultural evolution. It underlines the interaction of the environmental factor with the social dynamic. The model proposed herein is extremely widespread and involves the creation, maintenance, and diffusion of beliefs that afford a perspective on how such processes influence individual and collective psychological constructs. By synthesizing theories in the fields of biology, anthropology, and psychology, this research will argue that cultural evolution is one key mechanism able to explain both diversity and variability in human belief. The examination suggests that beliefs cannot be cast solely as some sort of passive variant of personal cognition; instead, they are actively generated by cultural dynamics and social learning techniques such as imitation and instruction. It is a book that places more emphasis on the role of political ideology in accepting scientific evidence and shows how directed reasoning leads to the perpetuation of noxious beliefs. It then investigates transfer biases supporting the reproduction of cultural traits across successive generations, hence affecting social structure and any effort of cooperation. This may lead to a nuanced grasp of how beliefs can be advantageous or disastrous, considering the prevailing social structures and ecology within which they live. Based on the wealth of research regarding under what circumstances beliefs arise and spread, this paper examines from a critical perspective psychological processes of belief systems and what they might portend for human behavior and social organization.

Kangaku and the State: Colonial Collaboration between Korean and Japanese Traditional Sinologists

This article examines the response of kangaku 漢学, or traditional sinological study, to modernity in Japan and colonial Korea by focusing on institutionalization and the interactions between scholars of traditional sinology in Japan and colonized Korea. In the Japanese Empire, kangaku actively demonstrated that it could contribute to the nation-state and became a part of the modern educational system, including the Imperial University. The favorable atmosphere toward kangaku in the Japanese Empire also enabled the institutionalization of kangaku in colonial Korea. The process of institutionalization there was deeply influenced by the traditional network of Japanese sinologists. Traditional sinologists in Japan and colonial Korea were able to cooperate based on a shared “shibun (斯文 “This Culture of Ours”) consciousness,” a kind of consciousness of crisis that originated in the Analects. The shibun consciousness was rooted in the traditional literary world, where borders and contemporaneity were relatively ambiguous. Within the university system, kangaku gradually faced the pressure of specialization and elaboration of modern scholarship and the demands of the nation-state. Thus, successive generations of Korean and Japanese sinologists moved toward a new modern ideology that went beyond “shibun consciousness.”

Resistance risk and mechanism of Ustilaginoidea virens to pydiflumetofen

Rice false smut, caused by Ustilaginoidea virens, is a devastating fungal disease in rice that not only leads to yield reduction but also poses a serious threat to food safety and human health due to the production of numerous mycotoxins. Pydiflumetofen, one of the most promising SDHI fungicides widely used for controlling various plant diseases, lacks available information regarding its antifungal activity against U. virens and the potential risk of resistance development in this pathogen. In this study, we evaluated the sensitivity of 33 field-isolated strains of U. virens to pydiflumetofen using mycelial growth inhibition method and assessed the potential for resistance development. The EC50 values for pydiflumetofen against the tested strains ranged from 0.0032 to 0.0123 μg/mL, with an average EC50 value of 0.0056 ± 0.0025 μg/mL. In addition, four strains of U. virens were randomly selected for chemical taming to evaluate their resistance risk to pydiflumetofen, resulting in the successful generation of eight stable and inheritable resistant mutants at a frequency of 1 %. These mutants exhibited significant differences in biological fitness compared to their respective parental strains. Cross-resistance tests revealed a correlation between pydiflumetofen and fluxapyroxad as well as fluopyram, but no evidence of cross-resistance was observed between pydiflumetofen and boscalid or tebuconazole. Therefore, we can conclude that the risk of resistance development in U. virens to pydiflumetofen is moderate. Finally, the target genes SDHB, SDHC, and SDHD in U. virens were initially identified, cloned, and sequenced to elucidate the mechanism underlying U. virens resistance to pydiflumetofen. Three mutation genotypes were found in the mutants: SDHB-H239Y, SDHB-H239L, and SDHC-A77V. The mutants carrying SDHB-H239Y exhibited low resistance, while SDHC-A77V showed moderate resistance, but the mutants with SDHB-H239L demonstrated high resistance. These findings contribute significantly to our comprehensive understanding of molecular mechanisms involved in the resistance of U. virens to pydiflumetofen, and provide an important reference for chemical control strategies against rice false smut in the field.

Antibody Modification via Lipoic Acid Ligase A-Mediated Site-Specific Labeling.

Enzymatic modification, particularly utilizing lipoic acid ligase (LplA), has emerged as a transformative approach in biopharmaceuticals, enabling precise and site-specific protein modifications. This review delves into the innovative applications of LplA in antibody modifications, including the creation of antibody-drug conjugates (ADCs) and the advancement of tag-free conjugation techniques. LplA's ability to facilitate the incorporation of bioorthogonal groups and its adaptability to various substrates underscores its versatility. Key developments include the successful generation of dual-labeled antibodies and the application of LplA in modifying antibody fragments. Additionally, the review explores the potential for LplA to enhance the therapeutic efficacy of ADCs through improved drug-to-antibody ratios and site-specific payload attachment. The implications of these advancements are significant, suggesting that LplA-mediated modifications could lead to more effective and targeted antibody-based therapies. This review aims to provide a comprehensive overview of LplA's role in expanding the possibilities of enzymatic conjugation, setting the stage for future research and clinical applications.

Frontiers in CAR-T Cell Therapy: Current Applications and Future Directions

CAR-T cell therapy, an ascending star in the immunotherapy landscape, harnesses the patient's innate immune defenses to confront cancer and a spectrum of hematologic disorders, while also exhibiting promising avenues for application in other maladies. This comprehensive review delves into the fundamental mechanisms of CAR-T cell therapy, the evolution and design strategies of successive generations of CAR-T cells, as well as the advancements and clinical implementations achieved in treating a diverse array of diseases.

Analysis of the winter oilseed rape recombination landscape suggests maternal-paternal bias.

Recombination, the reciprocal exchange of DNA between homologous chromosomes, is a mandatory step necessary for meiosis progression. Crossovers between homologous chromosomes generate new combinations of alleles and maintain genetic diversity. Due to genetic, epigenetic, and environmental factors, the recombination landscape is highly heterogeneous along the chromosomes and it also differs between populations and between sexes. Here, we investigated recombination characteristics across the 19 chromosomes of the model allopolyploid crop species oilseed rape (Brassica napus L.), using two unique multiparental populations derived from two genetically divergent founder pools, each of which comprised 50 genetically diverse founder accessions. A fully balanced, pairwise chain-crossing scheme was utilized to create each of the two populations. A total of 3213 individuals, spanning five successive generations, were genotyped using a 15K SNP array. We observed uneven distribution of recombination along chromosomes, with some genomic regions undergoing substantially more frequent recombination in both populations. In both populations, maternal recombination events were more frequent than paternal recombination. This study provides unique insight into the recombination landscape at chromosomal level and reveals a maternal-paternal bias for recombination number with implications for breeding.