High Homology Research Articles

Discovery of PF-07265028, A Selective Small Molecule Inhibitor of Hematopoietic Progenitor Kinase 1 (HPK1) for the Treatment of Cancer.

Hematopoietic progenitor kinase 1 (HPK1/MAP4K1) represents a high interest target for the treatment of cancer through an immune-mediated mechanism. Herein we present highlights of the drug discovery campaign within the lactam/azalactam series of inhibitors that yielded a small molecule (21, PF-07265028), which was advanced to a phase 1 clinical trial (NCT05233436). Key components of the discovery effort included optimization of potency through mitigation of ligand strain as guided by the use of cocrystal structures, mitigation of ADME liabilities (plasma instability and fraction metabolism by CYP2D6), and optimization of kinase selectivity, particularly over immune-modulating kinases with high homology to HPK1. Structure-based drug design via leveraging cocrystal structures and lipophilic efficiency analysis proved to be valuable tools that ultimately enabled the delivery of a clinical-quality small molecule inhibitor of HPK1.

Characterization and genomic analysis of the highly virulent Acinetobacter baumannii ST1791 strain dominating in Anhui, China.

The multidrug-resistant Acinetobacter baumannii clonal complex 92 is spreading worldwide due to its high-frequency gene mutation and recombination, posing a significant threat to global medical and health safety. Between November 2021 and April 2022, a total of 132 clinical A. baumannii isolates were collected from a tertiary hospital in China. Their growth ability and virulence of these isolates were assessed using growth curve analyses and the Galleria mellonella infection model. The genetic characteristics of the isolates were further examined through whole-genome sequencing. ST1791O/ST2P isolates represented the largest proportion of isolates in our collection and exhibited the highest growth rate and strongest virulence among all sequence types (STs) analyzed. Whole-genome sequences from 14,159 clinical isolates were collected from the National Center for Biotechnology Information database, and only nine ST1791O/ST2P isolates were detected. Comparative genomic analysis revealed that ST1791O/ST2P carried 11 unique genes, 5 of which were located within the capsular polysaccharide synthesis (cps) gene cluster. Single nucleotide polymorphisms (SNPs) between ST1791O/ST2P and other isolates were primarily found in the cps gene cluster. Among the other isolates, ST195O/ST2P and ST208O/ST2P exhibited the smallest SNP differences from ST1791O/ST2P, while ST195O/ST2P and ST1486O/ST2P had high homology. The ST1791O/ST2P strain in Anhui, China, displayed significant homology with ST195O/ST2P, ST208O/ST2P, and ST1486O/ST2P isolates. Compared to other isolates in this study, ST1791O/ST2P exhibited strong growth ability and virulence. Therefore, preventing the further spread of ST1791O/ST2P should be a top public health priority.

The path of GPR87: from a P2Y-like receptor to its role in cancer progression.

GPR87 is a G protein-coupled seven-transmembrane receptor first described as an orphan receptor in 2001. Despite its high structural homology to several extracellular nucleotide-activated P2Y receptors and sharing conserved sequence motifs in transmembrane regions, identification of endogenous ligands from the class of nucleotides and their analogues has failed for GPR87. Although lysophosphatidic acid was proposed to be a natural ligand for this cell surface receptor, these data are preliminary and inconsistent, and IUPHAR is currently considering GPR87 as an orphan receptor. Thus, the endogenous ligands and physiological functions of GPR87 are still required to be determined and/or confirmed. The remarkably higher expression of GPR87 in human malignant tissues compared to the normal healthy ones clearly suggests that this receptor may be involved in the development and progression of cancerous neoplasms. Therefore, in this review article, the main focus is placed on the oncogenic role of GPR87 in various human malignancies, presenting it as a potential novel target site for therapeutic interventions using both humanized monoclonal antibodies and gene therapy but also selective antagonists which are still waiting for their identification. Furthermore, the importance of the expression of GPR87 as a predictive biomarker for evaluating the prognosis and overall survival of cancer patients is also highlighted.

Genetic Profiling of Raoultella spp. in Pediatric Clinical Samples: Insights from Bauchi, North East Nigeria

Background: Raoultella species, are gram-negative bacteria of Enterobacteriaceae family, originally classified under Klebsiella, were reclassified in 2001 as a distinct genus. Members have emerged as notable pathogens, mostly in hospitalized and immunocompromised patients. Though once considered opportunistic, Raoultella spp. are now linked to various infections, including bacteremia, urinary tract infections, and neonatal sepsis, with antibiotic-resistant strains posing significant challenges in clinical settings. The true prevalence of Raoultella infections may be underreported due to diagnostic challenges, making early identification crucial for improved management in high-risk populations, particularly pediatrics. Raoultella species, emerging pathogens are increasingly linked to pediatric and neonatal infections in clinical settings. Aim: isolation and accurate identification of Raoultella species from pediatric clinical samples to address challenges in identification and genetic characterization. Study Design: This is a prospective, cross-sectional study involving pediatrics aged 0-5 years. Place and Duration of Study: Pioneering study in Bauchi, North-East Nigeria, from August 2021 to January 2022. Methodology: Samples from 262 pediatric patients with septicemia symptoms were processed at Abubakar Tafawa Balewa University Teaching Hospital (ATBUTH). The VITEK 2 Compact system was used for initial identification, while PCR amplification of the 16S rRNA gene confirmed Raoultella species. The genomic DNA of identified isolates underwent sequence and bioinformatics analysis. Results: The phylogenetic tree constructed using the 16S rRNA sequences of the isolates and reference strains visually represent the genetic relationships. The high homology observed in the sequence alignment is reflected in the phylogenetic clustering, with isolates RoP_2 (PQ213811), RoP_3 (PQ213812), and RoP_4 (PQ213813) grouping closely with known Raoultella planticola strains. This close genetic relationship further supports the species identification suggested by the gel electrophoresis and homology data. However, RoP_1 (PQ213810), identified as Proteus mirabilis through BLAST analysis of its 16S rRNA sequence, shows slightly lower homology with Raoultella planticola reference strains. In the phylogenetic tree, RoP_1 is positioned slightly apart from the other isolates, indicating some genetic variability. This discrepancy suggests that while RoP_1 shares characteristics with Raoultella, the distinct genetic makeup warrants further investigation to confirm its identification as R. planticola or otherwise. Conclusion: The phylogenetic analysis supports the identification of isolates RoP_2, RoP_3, and RoP_4 as Raoultella planticola, showing close genetic relationships. However, RoP_1 exhibits genetic variability, clustering separately and aligning more closely with Proteus mirabilis, suggesting the need for further investigation to confirm its species identification.

A pivotal switch in β-domain determines the substrate selectivity of TPS genes involved in Gardenia jasminoides floral scent synthesis.

Plants have evolved a diverse array of secondary metabolites to enhance their adaptability to environmental stresses, with volatile terpenoids being a notable example. Gardenia (Gardenia jasminoides), celebrated for its unique fragrance, is a key natural source of volatile terpenoids. Using our chromosome-level genome and transcriptome data for G. jasminoides, we previously identified six terpene synthases (TPS) involved in the production of its floral scent. Here, we functionally characterized these six key TPS enzymes, aligning their product profiles with volatile organic compound (VOC) analysis. Notably, we identified two highly similar terpene synthases, GjTPS1 and GjTPS2, which share high sequence homology but differ in substrate selectivity. By leveraging AI-based predictions and site-directed mutagenesis, we pinpointed a single amino acid in the β-domain that acts as a "switch," modulating substrate selectivity-an unusual finding, as this residue is outside the active site region. Comparative genomic analyses with Coffea canephora and other eudicots revealed that G. jasminoides TPS genes primarily expanded through dispersed (DSD) and tandem duplications (TD). Our study is the first to reveal a "switch" in a previously deemed "non-functional" region, regulating substrate selectivity in TPS genes. These insights could facilitate breeding elite gardenia varieties and support the rational engineering of terpenoid synthases.

Genome-wide survey of chlorophyllase (CLH) gene family in seven Rosaceae and functional characterization of MdCLH1 in apple (Malus domestica) leaf photosynthesis

Chlorophyll, a crucial pigment in plant photosynthesis, undergoes dynamic synthesis and degradation processes throughout the growth cycle of plants. Chlorophyllase (CLH) plays a crucial role in the degradation of chlorophyll by removing the phytol group from chlorophyll A to produce chlorophyllide A. However, Rosaceae species remain underexplored in terms of understanding the functional divergences among CLH gene family members (CLHs) involved in chlorophyll catabolism and photosynthesis. The apple (Malus domestica) CLHs also requires further systematic characterization and identification. In this study, 20 CLHs (MdCLH1-4, FvCLH1-2, PpCLH1-2, PcCLH1-3, PaCLH1-2, RrCLH1-4, RcCLH1-3) were identified from seven species belonging to the Rosaceae family. The chromosomal distribution of these gene family members is mostly separate across all species, except in Rosa rugosa. The number of amino acids encoded by these genes ranges from 171 aa to 391 aa, possessing a theoretical isoelectric point (PI) of 5.46-9.59, and a relative molecular weight of 18313.07D to 42413.21D. Secondary structure predictions highlight α-helix and random coil conformations as the dominant structural elements of CLH proteins present in Rosaceae species. Subcellular localization predictions indicate that all CLH proteins are expressed in chloroplasts, while MdCLH4 is uniquely localized to the nucleus. Phylogenetic analysis reveals high homology and close evolutionary relationships among the genes in three subfamilies. All these 20 CLHs contain elements responsive to phytohormones, environmental stress, and light. Furthermore, transcriptomic profiling using geneChip expression array coupled with qRT-PCR analyses revealed a heightened transcriptional activity of MdCLHs in leaf tissues and protective tissue of annual shoots as compared to other plant components. Additional experimental evidence specifically indicates MdCLH1 is located in the chloroplasts of tobacco leaves. Notably, MdCLH1 transient expression in apple leaves decreased chlorophyll a, carotenoids, total chlorophyll content, non - photochemical quenching coefficient (NPQ), and intercellular CO₂ concentration (Ci), while increasing chlorophyll b content, effective PSII quantum yield [Y(II)], net photosynthetic rate (Pn), stomatal conductance (Gs), and electron transport rate (ETR). These suggest MdCLH1 enhances light energy conversion in PSII by modulating chlorophyll degradation, potentially improving photosynthetic efficiency and reducing the potential for photoinhibition. This study lays a solid foundation for further exploration into the functional roles of CLHs in the Rosaceae family.

A multitask interpretable model with graph attention mechanism for activity prediction of low-data PIM inhibitors.

The aberrant expression of proviral integration site for Moloney murine leukemia virus (PIM) kinases is closely related to various tumors and chemotherapy resistance, making them attractive targets for cancer therapy. However, due to the extremely high homology among the three PIM isoforms (PIM1, PIM2, PIM3) and the limited availability of existing bioactivity data, screening and designing selective PIM inhibitors remain a daunting challenge. To address this issue, this study constructed a multitask regression model that can simultaneously predict the half-maximal inhibitory concentration (IC50 values). The model utilizes an attention mechanism to capture effects within local atomic groups and the interactions between different groups of atoms. Through weight sharing, the model enhances the accuracy of predicting PIM3 inhibitors by leveraging the rich and highly correlated data from PIM1 and PIM2 isoforms. Additionally, visualizing the weights of nodes (atoms in the molecule) in the model helps us to intuitively understand the relationship between molecular features and prediction outcomes, thereby enhancing the interpretability of the model. In summary, this work provides new insights and methods for performing activity prediction tasks for multiple similar targets in low-data scenarios.

Potential Virulence and Survival of Acid-Resistant Bacillus cereus KS100Bc Isolated from Foodservice Establishments in Korea

Bacillus cereus is an opportunistic pathogen causing food poisoning manifested by diarrheal or emetic syndromes. This study aimed to examine the potential virulence and survival of acid-resistant B. cereus strains isolated from foodservice establishments in schools. All isolates were positive for genes encoding nonhemolytic enterotoxin complexes, except for one strain that lacked nheC. More than 80% of the strains harbored enterotoxigenic genes. In contrast, cesB, which encodes an emetic toxin, was not detected. Among the isolates, the biofilm-forming activity of KS100Bc strain was 8.4 times higher than that of the type strain; therefore, KS100Bc was selected for further analyses. The KS100Bc survived in simulated gastric fluid (SGF) at pH 2.0 for 120 min, which is a major host barrier against exogenous microorganisms. The parameter Kmax of the log-linear + tail model for ATCC 14579 was up to1.31-fold higher than that for KS100Bc in SGF, indicating that it is difficult to inactivate KS100Bc in acidic gastric environments. Although whole-genome sequencing identified nucleotide sequences with high homology (>99%) between ATCC 14579 and KS100Bc, significant differences were observed in phenotypes such as biofilm formation and survival in SGF. The transcriptional profiles were compared to examine the differences in the response to acid shock. KS100Bc exhibited increased expression of genes associated with pH homeostasis, oxidative response, metabolic rearrangements, and general stress response with over twofold (P value < 0.05) relative gene expression compared with the type strain. These results suggest that identifying the characteristics of the acid-adapted isolate can broaden our understanding of therapeutic strategies and provide new directions for controlling and preventing the spread of an outbreak.

The Chicken HDAC4 Promoter and Its Regulation by MYC and HIF1A

Background: Histone deacetylase 4 (HDAC4) is a member of the class II histone deacetylase family, whose members play a crucial role in various biological processes. An in-depth investigation of the transcriptional characteristics of chicken HDAC4 can provide fundamental insights into its function. Methods: We examined HDAC4 expression in chicken embryonic stem cells (ESC) and spermatogonial stem cells (SSC) and cloned a 444 bp fragment from upstream of the chicken HDAC4 transcription start site. Subsequently, we constructed pEGFP-HDAC4 and a series of 5′-deletion luciferase reporter constructs, which we transfected into DF-1 cells to measure their transcriptional activity. The regulatory mechanisms of chicken HDAC4 expression were investigated by performing trichostatin A (TSA) treatment, deleting putative transcription factor binding sites, and altering transcription factor expression levels. Results: HDAC4 exhibited higher expression in SSC than in ESC. We confirmed that the upstream region from −295 bp to 0 bp is the core transcriptional region of HDAC4. TSA effectively inhibited HDAC4 transcription, and bioinformatics analysis indicated that the chicken core HDAC4 promoter sequence exhibits high homology with those of other avian species. The myelocytomatosis viral oncogene homolog (MYC) and hypoxia-inducible factor 1 α (HIF1A) transcription factors were predicted to bind to this core region. Treatment with TSA for 24 h resulted in the upregulation of MYC and HIF1A, which repressed HDAC4 transcription. Conclusions: Our results provide a basis for subsequent investigations into the regulation of HDAC4 expression and biological function.

Recent Trends in Development of Novel Therapeutics for Modulation of 14-3-3 Protein-Protein Interactions in Diseases.

14-3-3s constitute a group of proteins belonging to the phosphoserine/phosphothreonine family that are involved in the regulation of several physiological pathways by interacting with several client proteins. All the eukaryotic cells are known to possess 14-3-3 isoforms. In addition, 14-3-3s isolated from different eukaryotic cells share high sequence homology with each other. Seven isoforms (β, γ, ε, η, ζ, σ, and τ/θ) have been yet identified in mammals. These proteins participate in several physiological processes by either stimulating or interfering with the enzymatic activities of binding partners. These proteins take part in several human diseases upon dysregulation which include cancer and neurodegenerative disorders. Recently, a number of evidences suggest that the interaction of 14-3-3s with either oncogenic or pro-apoptotic proteins can lead to cancer development in animals. In the case of neurodegenerative disorders, 14-3-3s interact with Lewy bodies and neurofibrillary tangles in Parkinson's and Alzheimer's diseases, respectively. The current review focuses on strategies to regulate 14-3-3s' proteins in diseases. Potential strategies to regulate 14-3-3 interactions in disease conditions include the use of small interfering RNAs (siRNA), microRNA (miRNA), and modifications of 14-3-3s or their client proteins. In addition, some peptides or chemicals can also serve as potential inhibitors of 14-3-3. However, optimization of these therapeutic strategies is required for their practical implementations.

Bioinformatics, expression, purification, and inflammation-inducing effect of Mycoplasma genitalium GroEL protein

To preliminarily understand the pathogenic mechanism of Mycoplasma genitalium (Mg) GroEL protein, we used bioinformatics tools to predict the structure and function of Mg GroEL protein and then constructed the recombinant plasmid pET-28a-GroEL. The protein expression was induced by 0.2 mmol/L IPTG, and the expressed protein was purified by Ni-iminodicitic acid (IDA) column affinity. Tohoku Hospital Pediatrics-1 (THP-1) cells were exposed to 2 μg/mL Mg rGroEL. The levels of interleukin (IL)-1β and tumor necrosis factor (TNF)-α in the cell supernatant were measured by ELISA, and that of IL-6 was measured by an automatic chemiluminescence instrument. The activation of the nuclear factor-kappa B (NF-κB) signaling pathway was visualized by immunofluorescence and Western blotting. The results showed that Mg GroEL was a stable hydrophilic protein composed of 543 amino acid residues, with the relative molecular mass of 58.44 kDa, an isoelectric point of 5.68, and a molecular formula of C2568H4300N700O825S8. The secondary structure was mainly composed of α-helices and random coils. Mg GroEL contained 12 B-cell dominant epitopes and 10 T-cell dominant epitopes. It exhibited high homology with the GroEL proteins from Mycoplasma pneumoniae, M. agalactiae, M. arthritidis, M. hyopneumoniae, and M. bovis. Mg rGroEL activated the NF-κB signaling pathway and promoted the secretion of IL-1β, IL-6, and TNF-α in THP-1 cells. These results suggest that Mg GroEL exhibits substantial antigenicity and possesses the capability of triggering inflammation in host cells. This study establishes a theoretical basis for future investigations pertaining to the role and pathogenic mechanisms of Mg GroEL.

Identification and functional analysis of sex-determining genes in the spongy moth, Lymantria dispar (lepidoptera: Erebidae)

The spongy moth (Lymantria dispar) employs a female heterogametic sex-determination system, where the female sex-determining factor (F factor) is located on the W chromosome, and the male sex-determining factor (M factor) is located on the Z chromosome. The sex-determining capabilities of the F factor and M factor vary among subspecies. Consequently, L. dispar serves as an excellent model for studying the mechanisms underlying the evolution and diversity of sex-determining genes. However, the genes encoding the F and M factors, as well as the molecular functions of their translation products, remain unidentified.In this study, we identified a L. dispar Masculinizer ortholog (LdMasc) and found that this gene is highly expressed in male embryos during the sex-determination stage. When LdMasc expression was silenced using embryonic RNA interference (RNAi), the expression pattern of L. dispar doublesex (Lddsx), the master regulatory gene for sex differentiation, shifted from the male-specific form to the female-specific form in male embryos. To identify potential F factors, we screened for genes that were exclusively expressed in females across multiple tissues and located only within the female genome. This screening yielded four unigenes with sequences displaying high homology to each other. These unigenes formed a tandem repeat, comprising approximately 100 copies within a 200 kbp region of the W chromosome-derived contig. We designated these unigenes as Fet-W (female-specifically expressed transcript from the W chromosome). RT-PCR analysis revealed that Fet-W was expressed in a female-specific manner during the sex-determination stage. Suppression of Fet-W expression by embryonic RNAi led to an increase in LdMasc expression in females and a corresponding shift in dsx expression patterns from the female-specific to the male-specific form. These findings strongly suggest that the F factor in L. dispar is Fet-W, whereas the M factor is LdMasc.

Genetic Characterization of a Novel Retron Element Isolated from Vibrio mimicus.

Bacterial reverse transcriptase coding gene (RT) is essential for the production of a small satellite DNA-RNA complex called multicopy single-stranded DNA (msDNA). In this study, we found a novel retron, retron-Vmi1 (Vm85) from Vibrio mimicus. The retron is comprised of the msr-msd region, orf323, and the ret gene, a genetic organization similar to Salmonella's retron-Sen2 (St85). The protein sequence of the RNA-directed DNA polymerase (RT-Vmi1) is highly homologous to the RTs of Vibrio metoecus, Vibrio parahaemolyticus, and Vibrio vulnificus. Phylogenetic and protein sequence similarity analysis of retron-Vmi1 ORF323 and RT revealed a close relatedness to retron-Sen2. We found that retron-Vmi1 was inserted in the dusA gene, similar to the insertion of the retron-Vpa1 (Vp96) of V. parahaemolyticus AQ3354, suggesting that retrons can be transferred via the tRNA gene. These results are the first convincing evidence that retron is moving across species. The neighboring genes of retron-Vmi1 shared high homology with the genetic environment of V. parahaemolyticus and V. vulnificus retrons. We also found two junction points within the retron-Vmi1 and the dusA gene suggesting that retron-Vmi1 was inserted into this site in a two-step manner.

Concurrent Coxsackievirus A6 Infection and Kawasaki Disease: A Case Report

Background and Clinical Significance: Kawasaki disease (KD) is an acute febrile vasculitis that primarily affects children and is associated with systemic inflammation, particularly in the coronary arteries. Coxsackievirus A6 (CVA6) has emerged as a significant agent in atypical presentations of hand, foot, and mouth disease (HFMD), raising the possibility of its involvement in KD. Case Presentation: This report presents the case of an 18-month-old Thai boy admitted with symptoms of high fever, sore throat, and ulcerative lesions, initially diagnosed with herpangina. As his condition progressed, additional KD symptoms developed, including conjunctival injection, rash, and elevated inflammatory markers, fulfilling the diagnostic criteria for KD. Notably, throat swab analysis confirmed CVA6 as the causative agent. Phylogenetic analysis revealed that the CVA6 strain closely aligned with Chinese strains from 2023, showing a high nucleotide sequence homology of 98.4%. Conclusions: In conclusion, this case highlights a possible association between CVA6-associated herpangina and KD, suggesting that CVA6 infection may act as a trigger for KD in genetically susceptible children. These findings highlight the need for increased awareness among healthcare providers to promptly identify and manage Kawasaki Disease during peak enterovirus seasons, reducing its impact on children.

Genome-wide identification, transcript profiling and functional analyses of PCP gene family in Wucai (Brassica campestris)

Pollen coat proteins (PCPs) are cysteine-rich small-molecule proteins, which exhibit high levels of polymorphism and are expressed in gametocytes. Previous investigations have revealed that PCP genes are involved in pollen wall synthesis, pollen-stigma recognition, and pollen development and germination. However, gene expression and function of PCP family in pollen development is not well understood in Wucai (Brassica campestris L.). In this study, genome-wide identification and expression analysis of the BcPCP gene family members were conducted, including their physical and chemical properties, chromosome localization, phylogenetic relationships, gene structure, and tertiary structure. A total of 20 BcPCP genes were identified and classified into three subfamilies showing high homology to Arabidopsis thaliana. Expression pattern analysis indicated that the BcPCP gene family exhibits higher expression levels in reproductive organs, suggesting their potential involvement in the reproductive development. Notably, BraA02g002400.3 C, potentially associated with male sterility, was identified through multiple transcriptomic and proteomic datasets. Subsequently, sequence analysis revealed its homology with the Arabidopsis GRP20 gene, and thus it was named BcGRP20. Functional analysis of this gene showed that overexpression of BcGRP20 gene in the Arabidopsis grp20 mutant could restore anther fertility. Overall, our findings indicate that BcGRP20 plays a critical role in pollen development and may be the causative gene for male sterility in Wucai. This study provides candidate genes for further functional identification of BcPCP genes in Wucai, which are crucial for anther development.

Genome-Wide Identification of Heat Shock Transcription Factor Family and Key Members Response Analysis to Heat Stress in Loquat

Eriobotrya japonica (loquat) is an evergreen fruit tree of the apple tribe in Rosaceae with high edible and medicinal value. The yield and quality of loquat fruits are significantly influenced by environmental stress, particularly heat stress during fruit ripening. In this study, thirty EjHSFs were identified in the loquat genome. Twenty-nine EjHSFs were unevenly distributed across sixteen chromosomes, except Chr-6. A synteny analysis revealed that twenty-six EjHSF genes had undergone duplication events. Twenty-nine EjHSF genes were found to be in sync with HSF in apples while also diverging with other Rosaceae species. A phylogenetic analysis revealed that EjHSFs could be divided into three categories, including eighteen EjHSF-A, ten EjHSF-B, and two EjHSF-C. Twenty-nine members of the EjHSF family showed high homology to those of Malus domestica and Gillenia trifoliate. A promoter analysis retrieved thirty-three CAREs that were functionally relevant and connected to the expression of HSFs. Among these, the promoters of twenty-three EjHSF genes possessed at least one STRE element that could be activated by heat shock. Eleven of these EjHSFs were highly expressed in loquat tissues, namely EjHSF-B4a, EjHSF-A4a, EjHSF-A4d, and EjHSF-C1a in roots; EjHSF-B4b in roots and inflorescence; EjHSF-C1b in stems and roots; EjHSF-A2a in three tissues; EjHSF-A8b in four tissues; and EjHSF-A4c, EjHSF-B1a, and EjHSF-B2b in six tissues. Moreover, fifteen EjHSFs were differentially expressed (eleven upregulated and four downregulated) in fruits under heat stress treatment in the color-changing period. Among them, EjHSF-A2a and EjHSF-A2b upregulated transcriptional abundance over 300 times after heat treatment; EjHSF-B2b consistently displayed an increasing trend over time; and EjHSF-B1a was significantly downregulated. Hence, these results suggest that EjHSFs take part in loquat tissue development and may be involved in the fruit’s heat stress response. These findings enhance the understanding of EjHSFs’ role within loquats and the heat stress response of its fruit and provide target genes for heat stress improvement.

Charting Cortical-Layer Specific Area Boundaries Using Gibbs Ringing Attenuated T1w/T2w-FLAIR Myelin MRI.

Cortical areas have traditionally been defined by their distinctive layer cyto- and/or myelo- architecture using postmortem histology. Recent studies have delineated many areas by measuring overall cortical myelin content and its spatial gradients using the T1w/T2w ratio MRI in living primates, including humans. While T1w/T2w studies of areal transitions might benefit from using the layer profile of this myelin-related contrast, a significant confound is Gibbs ringing artefact, which produces signal fluctuations resembling cortical layers. Here, we address these issues with a novel approach using cortical layer thickness-adjusted T1w/T2w-FLAIR imaging, which effectively cancels out Gibbs ringing artefacts while enhancing intra-cortical myelin contrast. Whole-brain MRI measures were mapped onto twelve equivolumetric layers, and layer-specific sharp myeloarchitectonic transitions were identified using spatial gradients resulting in a putative 182 area/subarea partition of the macaque cerebral cortex. The myelin maps exhibit unexpectedly high homology with humans suggesting cortical myelin shares the same developmental program across the species. Comparison with histological Gallyas myelin stains explains over 80% of the variance in the laminar T1w/T2w-FLAIR profiles, substantiating the validity of the method. Altogether, our approach provides a novel, noninvasive means for precision mapping layer myeloarchitecture in the primate cerebral cortex, advancing the pioneering work of classical neuroanatomists.

First Report of Rhizopus arrhizusi causing rot of Dictyophora rubrovolvata in China.

Dictyophora rubrovolvata, as an edible fungus with high medicinal value, is widely cultivated in several provinces in China (Hang et al. 2012). However, between December 2023 and March 2024, a rot disease occurred in the main production area in Fengxian District, Shanghai, China (N30°93', E121°49'). The disease incidence was 25% in the affected 1.33-ha growing area. High temperatures (>25℃) and poor ventilation provide favorable conditions for the spread of this disease. The disease mainly occurs at the stage of fruiting bodies formation of D. rubrovolvata. When the epidermis is damaged and broken, it becomes infested with mold, which then produces a layer of moldy rot with pus. The infected D. rubrovolvata tissues at the edge of the lesions were isolated, surface sterilized and cultured on potato dextrose agar (PDA) at 30 ℃ under dark conditions. Pure cultures were obtained by single-spore isolation. After 3 days, isolates were transferred to Czapek Yeast agar (CYA) (Samson et al, 2014). On CYA, the fungal colony consisted of white flocculent hyphae. Scanning electron microscopy analysis showed that the mycelium was white, and the internodes of the stolons formed characteristic pseudoroots, from which upwardly clustered erect, unbranched sporocarp peduncles expanded apically to form rounded sporocarp sacs, within which sporocarp spores were produced. (Hariprasath P, 2019). To confirm the identity of the pathogen, the genomic fragments for the internal transcribed spacer (ITS) and intergenic spacer (IGS) gene of the isolate were amplified by PCR (White et al. 1990; Liu XY. 2008). The resulting sequence was deposited in GenBank with accession PP951880 and PQ001670, respectively. PCR results and morphological observations indicated the isolated strain was a pure culture and the strain was designated as DIC01. Comparative results showed that the sequences with accession numbers MT603964.1 and DQ990323.1 showed high homology of 99.15% and 98.96% to the ITS and IGS sequences of Rhizopus arrhizusi DIC01, respectively. Phylogenetic analysis with ITS and IGS genes of the isolated strain and 7 Rhizopus spp. strains were performed using MEGAX with Neighbor-Joining (NJ) method. Based on the results of growth habits, morphological observations, and phylogenetic analysis, the pathogen was identified as R. arrhizusi. A spore suspension of the R. arrhizusi DIC01 (1 x107 conidia/mL) was inoculated back to healthy D. rubrovolvata. Five healthy fruit bodies of D. rubrovolvata were injected, and another five healthy morels were treated with potato dextrose broth (PDB) as controls. D. rubrovolvata was incubated at 25°C and 90% relative humidity without ventilation for 5 days. The pathogen successfully infected the D. rubrovolvata, which developed white moldy lesions similar to those of natural diseases. The controls remained healthy without any symptoms. The pathogen was reisolated from the affected lesions and identified as R. arrhizusi DIC01 based on its morphological characteristics and phylogenetic marker genes. R. arrhizusi has been reported to cause endothelial cell damage and mycelial invasion into blood vessels, leading to thrombosis and tissue necrosis. (Hariprasath P, 2019). To our knowledge, this is the first report of R. arrhizusi causing rot disease of D. rubrovolvata. This study confirmed that R. arrhizusi is the pathogenic fungus responsible for rotting disease in D. rubrovolvata farms in Fengxian, Shanghai.

Dengue Virus Infection: Immune Response and Therapeutic Targets.

Flavivirus infection, especially dengue virus infection caused by DENV, is known to be a significant health concern globally owing to the high incidence and mortality rate. The expanding and increasing disease burden calls for the need to develop an effective treatment and prevent the event of fatal complications, including dengue hemorrhagic fever/dengue shock syndrome. The DENV-induced immune response has been described as paradoxical because it has a protective role in viral clearance but, at the same time, causes more severe infection through viral-specific immunity. This is further complicated by high homology and cross-reactivity between different serotypes of DENV, causing a more severe disease presentation during secondary infection by a heterologous serotype. This serotype complexity poses a challenge for the development of a universal flavivirus vaccine. This review highlights the significance of high motility group box 1 (HMGB1) and nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing 3 (NLRP3) inflammasome activation pathways in initiating an inflammatory response through the downstream activation of nuclear factor κB and proinflammatory cytokine Interleukin (IL)-1B, IL-18 release in DENV infection. It also discusses the role of NLRP3 in activating cellular apoptosis and pyroptosis leading to systemic failure, especially in peripheral tissues. Over the decades, there has been much progress in understanding the immunopathogenesis of DENV infection. Researchers have been studying key pathogenic molecules for potential therapeutic targets including HMGB1 and NLRP3 inflammasome inhibitors, which is explored in this review. Ultimately, although there is not yet an effective antiviral or vaccine for DENV, immunomodulators continue to pave the way to decrease disease severity in infected individuals.

FNDC4 alleviates cardiac ischemia/reperfusion injury through facilitating HIF1α-dependent cardiomyocyte survival and angiogenesis in male mice

Fibronectin type III domain-containing (FNDC) proteins play critical roles in cellular homeostasis and cardiac injury, and our recent findings define FNDC5 as a promising cardioprotectant against doxorubicin- and aging-related cardiac injury. FNDC4 displays a high homology with FNDC5; however, its role and mechanism in cardiac ischemia/reperfusion (I/R) injury remain elusive. Here, we show that cardiac and plasma FNDC4 levels are elevated during I/R injury in a hypoxia-inducible factor 1α (HIF1α)-dependent manner. Cardiac-specific FNDC4 overexpression facilitates, while cardiac-specific FNDC4 knockdown inhibits cardiomyocyte survival and angiogenesis in I/R-stressed hearts of male mice through regulating the proteasomal degradation of HIF1α. Interestingly, FNDC4 does not directly stimulate angiogenesis of endothelial cells, but increases the expression and secretion of fibroblast growth factor 1 from cardiomyocytes to enhance angiogenesis in a paracrine manner. Moreover, therapeutic administration of recombinant FNDC4 protein is sufficient to alleviate cardiac I/R injury in male mice, without resulting in significant side effects. In this work, we reveal that FNDC4 alleviates cardiac I/R injury through facilitating HIF1α-dependent cardiomyocyte survival and angiogenesis, and define FNDC4 as a promising predictive and therapeutic target of cardiac I/R injury.