Modulation Of Growth Research Articles

Abstract 7235: Exosome-mediated delivery of siRNA against NRF2 for treatment of lung cancer

Abstract Nuclear factor erythroid 2-related factor 2 (NRF2) activation is the third most common genetic event in lung adenocarcinomas (25%), primarily induced by loss of function mutations in its master regulator, KEAP1. NRF2 is a key modulator of tumor growth and metabolic reprogramming, resulting in resistance to traditional chemotherapeutics. Due to its role in modulating oxidative stress, sustained NRF2 overexpression promotes disease progression, particularly in lung cancer. This study demonstrates the effective knockdown of NRF2 expression by small interfering RNA (siRNA) delivered via a nanoplatform based on bovine colostrum exosomes to lung cancer cells, both in vitro and in vivo, resulting in inhibition of proliferation and disease progression. Bovine colostrum is an abundant source of exosomes. Here we present a novel exosome-polyethyleneimine matrix (EPM) to deliver therapeutic siRNA targeting NRF2 expression (siNRF2). Specific delivery of the EPM to tumors was further enhanced via folic acid (FA) functionalization, targeting folate receptors highly expressed on lung tumor cells. Exosomes were isolated from standardized colostrum powder through rehydration and differential centrifugation, and characterized by size, polydispersity index (pdi) and surface charge by Zetasizer, and the presence of surface protein markers by Western blot. siNRF2 was loaded onto the EPM, and the resultant EPM-siNRF2 complex was harvested by PEG precipitation. Entrapment efficiency was determined using 5’-32P-labeled siRNA as a tracer. The gene knockdown efficiency of EPM-siNRF2 was assessed in lung cancer (A549) cells by Western blot and the therapeutic effect of target knockdown in vitro was confirmed by analyzing the corresponding modulation in downstream targets of NRF2. The anti-tumor effect was then evaluated using both subcutaneous and orthotropic lung tumor models in immunocompromised mice. The EPM exhibited high entrapment efficiency (>90%) of siNRF2 (up to 20 μg) and protected entrapped siRNA from enzymatic degradation upon exposure to RNases. EPM-siNRF2 dose dependently decreased NRF2 expression in A549 lung cancer cells. When formulated with FA-functionalized exosomes, EPM-siNRF2 inhibited subcutaneous and orthotropic lung tumors in mice by over 57% (p<0.01) and 84% (p<0.001), respectively. Mice treated with FA-EPM-siNRF2 showed decreased levels of NRF2 in tumor tissue, but not in the adjacent lung, indicating FA-mediated targeted delivery. This bovine colostrum exosome based EPM technology showcases high efficiency in gene knockdown and lung tumor inhibition, providing a promising nanoplatform for delivering nucleic acid therapeutics to the tumor site. Funding: Supported from the NIH grant R44 CA-221487, 3P Biotechnology Contract and, in part, Agnes Brown Duggan Endowment. Keywords: Bovine colostrum, Exosomes, NRF2, siRNA, Lung cancer. Citation Format: Raghuram Kandimalla, Margaret Wallen, Jeyaprakash Jeyabalan, Disha N. Moholkar, Wendy Spencer, Ramesh C. Gupta, Farrukh Aqil. Exosome-mediated delivery of siRNA against NRF2 for treatment of lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7235.

Abstract 3085: Cholesterol sulfotransferase SULT2B1b suppresses prostate tumor growth and regulates cholesterol metabolism in prostate cancer cells

Abstract Prostate cancer (PrCa) ranks as the second leading cause of cancer death in males in the United States. The 5-year survival rate drops drastically from 100% in patients with early-stage confined PrCa to 30% in those with advanced metastatic PrCa. Patients with advanced PrCa face limited long-term treatment options, often developing resistance to androgen-deprivation therapy and chemotherapy. Identifying and targeting pivotal molecules or pathways in advanced PrCa can substantially enhance patient survival. Our previous findings unveiled an inverse relationship between cholesterol sulfotransferase SULT2B1b and the severity of human PrCa. The highest levels of SULT2B1b were observed in the epithelium of normal prostates, with a progressive decrease as cancer advanced. The lowest levels were found in the most advanced metastatic samples. This study aims to determine whether SULT2B1b performs any active and critical functions in PrCa progression and assess whether it regulates cholesterol metabolism in PrCa cells. We generated SULT2B1b knockout (KO) clones from mouse PrCa cell line MyC-CaP and established inducible SULT2B1b overexpression lines from androgen receptor (AR)+ LNCaP and AR- PC3 human PrCa cells. These modified mouse and human cells were implanted into syngeneic FVB mice and immunodeficient NRG mice, respectively. MyC-CaP KO cells formed significantly larger tumors compared to parental cells, accompanied by reduced survival rates. In contrast, both LNCaP and PC3 tumors with induced SULT2B1b exhibited smaller sizes compared to control tumors, leading to improved survival rates. To investigate the impact of SULT2B1b on cholesterol metabolism and the underlying mechanisms behind SULT2B1b-regulated tumor growth, we examined genes related to lipoprotein signaling and cholesterol metabolism in MyC-CaP SULT2B1b KO cells using PCR arrays and qRT-PCR. Genes responsible for cholesterol uptake and synthesis (e.g. Ldlr, Cyp51, Sorl1, Pcsk9 and Insig1) were among the top upregulated genes, while genes controlling cholesterol efflux (e.g. Abca1) and esterification (e.g. Soat1 and Soat2) were downregulated in KO cells. Consistent with these gene expression patterns, intracellular cholesterol levels significantly increased in KO cells. Interestingly, although SULT2B1b did not affect cell proliferation under normal conditions, KO cells exhibited faster growth than parental cells when cultured in lipid-depleted or charcoal-stripped media, possibly due to altered intracellular cholesterol levels. In summary, these findings align with the expression pattern of SULT2B1b in human PrCa, demonstrating that SULT2B1b inhibits tumor growth, whereas its removal promotes tumor growth. This suggests SULT2B1b may serve as a critical tumor suppressor in PrCa. Its modulation of tumor growth may occur through the regulation of cholesterol metabolism. Citation Format: Jiang Yang, Dilinaer Wusiman, Timothy L. Ratliff. Cholesterol sulfotransferase SULT2B1b suppresses prostate tumor growth and regulates cholesterol metabolism in prostate cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3085.

Abstract 2558: Molecularly matched targeted therapies benefit high risk pediatric cancer patients

Abstract Introduction: Genomic technologies have the potential to transform the paradigm for diagnosing and treating children with cancer. Although druggable alterations are identified in a large fraction of pediatric cancer patients, the extent to which point-of-care genomic testing directly impacts therapeutic outcomes of individual patients in real-world settings is less well-known. A review of patients sequenced through the Precision in Pediatric Sequencing (PIPseq) program at Columbia University Irving Medical Center (CUIMC) was conducted to describe outcomes of patients treated with a molecularly matched targeted therapy (MTT). Methods: Matched tumor-normal whole exome and tumor transcriptome sequencing was conducted in a CLIA-certified laboratory at CUIMC between January 2013 and March 2023 on samples from 373 high risk pediatric cancer patients including those with <50% survival, rare cancers lacking standard therapies, and relapsed/refractory disease. All cases were reviewed at a multi-disciplinary molecular tumor board and reports were transmitted to the electronic health record (EHR). EHR data were reviewed to identify patients who received a MTT. Tumor-specific response and progression-free survival (PFS) among the MTT cohort were investigated. Clinical benefit was further assessed using the Growth Modulation Index (GMI) whereby patients served as their own control. GMI is the ratio of PFS on the MTT therapy to the PFS on the most recent prior therapy. A ratio of >/=1.3 indicates meaningful clinical activity. Results: Thirty pediatric patients (solid tumors, n=5; CNS tumors, n=15; hematologic malignancies, n=10) received a MTT. Median age at sequencing was 11 years, range 10 months-23 years. Median time to MTT from sequencing was 4.5 months, range 7 days to 19.2 months. Best overall response included 7 complete responses (23%), 6 partial responses (20%), 5 with stable disease (17%), 10 with progressive disease (33%), and 2 indeterminate due to early death or lack of follow-up during therapy (7%). GMI was calculated for 19/30 patients (63%); median GMI=1.1. 11 patients were excluded due to lack of prior therapy, early death, insufficient follow-up, or lack of progression between prior and targeted therapies. GMI ratios indicated clinical benefit from MTT for 12/19 patients (63%), median GMI=1.5. Conclusions: Clinical comprehensive molecular profiling for pediatric patients with high risk cancers identified patients who are candidates for MTT. Overall, MTT led to stable disease, partial or complete response, for 60% of patients. Utilizing GMI ratios as an indicator of clinical benefit for select patients provided additional information beyond traditional endpoints. Twelve patients with GMI >/=1.3 experienced improved PFS compared to their most recent prior therapy. These results demonstrate real-world clinical benefit from MTT, supporting molecular profiling of pediatric high risk tumors. Citation Format: Alice Tao, Jovana Pavisic, Jennifer A. Oberg. Molecularly matched targeted therapies benefit high risk pediatric cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2558.

The Effect of Heparin and Other Exogenous Glycosaminoglycans (GAGs) in Reducing IL-1β-Induced Pro-Inflammatory Cytokine IL-8 and IL-6 mRNA Expression and the Potential Role for Reducing Inflammation.

Glycosaminoglycans (GAGs) are long linear polysaccharides found in every mammalian tissue. Previously thought only to be involved in cellular structure or hydration, GAGs are now known to be involved in cell signaling and protein modulation in cellular adhesion, growth, proliferation, and anti-coagulation. In this study, we showed that GAGs have an inhibitory effect on the IL-1β-stimulated mRNA expression of IL-6 and IL-8. Exogenous heparin (p < 0.0001), heparan (p < 0.0001), chondroitin (p < 0.049), dermatan (p < 0.0027), and hyaluronan (p < 0.0005) significantly reduced the IL-1β-induced IL-8 mRNA expression in HeLa cells. Exogenous heparin (p < 0.0001), heparan (p < 0.0001), and dermatan (p < 0.0027) also significantly reduced IL-1β-induced IL-6 mRNA expression in HeLa cells, but exogenous chondroitin and hyaluronan had no significant effect. The exogenous GAGs may reduce the transcription of these inflammatory cytokines through binding to TILRR, a co-receptor of IL-1R1, and block/reduce the interactions of TILRR with IL-1R1.

Dynamic Modulation of Li2O2 Growth in Li-O2 Batteries through Regulating Oxygen Reduction Kinetics with Photo-Assisted Cathodes.

Widely acknowledged that the capacity of Li-O2 batteries (LOBs) should be strongly determined by growth behaviors of the discharge product of lithium peroxide (Li2O2) that follows both coexisting surface and solution pathways. However until now, it remains still challenging to achieve dynamic modulation on Li2O2 morphologies. Herein, the photo-responsive Au nanoparticles (NPs) supported on reduced oxide graphene (Au/rGO) have been utilized as cathode to manipulate oxygen reduction reaction (ORR) kinetics by aid of surface plasmon resonance (SPR) effects. Thus, we can experimentally reveal the importance of matching ORR kinetics with Li+ migration towards battery performance. Moreover, it is found that Li+ concentration polarization caused "sudden death" of LOBs is supposed to be just a form of suspended animation that could timely recover under irradiation. This work provides us an in-depth explanation on the working mechanism of LOBs from a kinetic perspective, offering valuable insights for the future battery design.

Modulation of growth, microcystin production, and algal-bacterial interactions of the bloom-forming algae Microcystis aeruginosa by a novel bacterium recovered from its phycosphere.

Harmful algal blooms (HABs) in natural waters are of escalating global concern due to their detrimental impact on environmental health. Emerging evidence indicates that algae-bacteria symbionts can affect HAB features, though much about this interplay remains largely unexplored. The current study isolated a new species of Mucilaginibacter (type strain JXJ CY 39T) from culture biomass of the bloom-causing Microcystis aeruginosa FACHB-905 (Maf) from Lake Dianchi, China. Strain JXJ CY 39T was an aerobic, Gram-stain-negative rod bacterium that grew at 5-38°C, pH 4.0-11.0, and 0-3.0% NaCl. Taxonomic evaluation proposed a new species, with Mucilaginibacter lacusdianchii sp. nov., as the species epithet. Experimental results revealed that strain JXJ CY 39T spurred the growth of Maf by supplying soluble phosphorus and nitrogen during cultivation, despite the unavailability of soluble phosphorus and nitrogen. Additionally, by producing the plant hormone indole-3-acetate, strain JXJ CY 39T possibly impacted Maf's functionality. Results from co-culture experiments with other strains from Maf biomass showed possible effects of strain JXJ CY 39T on the relationship between Maf and other cohabiting bacteria, as well as microcystin toxin production characteristics. Although Maf could foster the growth of strain JXJ CY 39T by supplying organic carbon, the strain's growth could be regulated via specific chemical compounds based on antibiotic assays. Community composition analysis disclosed that this Mucilaginibacter strain positively affected Maf's growth and modified densities and types of bacteria linked to Maf. Overall, these results suggest that the interactions between important HAB-causing organisms and their attached bacteria are complex, dynamic, and may influence the growth characteristics of algae.

Automated peripancreatic vessel segmentation and labeling based on iterative trunk growth and weakly supervised mechanism

Peripancreatic vessel segmentation and anatomical labeling are pivotal aspects in aiding surgical planning and prognosis for patients with pancreatic tumors. Nevertheless, prevailing techniques often fall short in achieving satisfactory segmentation performance for the peripancreatic vein (PPV), leading to predictions characterized by poor integrity and connectivity. Besides, unsupervised labeling algorithms usually cannot deal with complex anatomical variation while fully supervised methods require a large number of voxel-wise annotations for training, which is very labor-intensive and time-consuming. To address these two problems, we propose an Automated Peripancreatic vEssel Segmentation and lAbeling (APESA) framework, to not only highly improve the segmentation performance for PPV, but also efficiently identify the peripancreatic artery (PPA) branches. There are two core modules in our proposed APESA framework: iterative trunk growth module (ITGM) for vein segmentation and weakly supervised labeling mechanism (WSLM) for artery labeling. The ITGM is composed of a series of iterative submodules, each of which chooses the largest connected component of the previous PPV segmentation as the trunk of a tree structure, seeks for the potential missing branches around the trunk by our designed branch proposal network, and facilitates trunk growth under the connectivity constraint. The WSLM incorporates the rule-based pseudo label generation with less expert participation, an anatomical labeling network to learn the branch distribution voxel by voxel, and adaptive radius-based postprocessing to refine the branch structures of the labeling predictions. Our achieved Dice of 94.01% for PPV segmentation on our collected dataset represents an approximately 10% accuracy improvement compared to state-of-the-art methods. Additionally, we attained a Dice of 97.01% for PPA segmentation and competitive labeling performance for PPA labeling compared to prior works. Our source codes will be publicly available at https://github.com/ZouLiwen-1999/APESA.

A natural variation in OsDSK2a modulates plant growth and salt tolerance through phosphorylation by SnRK1A in rice.

Plants grow rapidly for maximal production under optimal conditions; however, they adopt a slower growth strategy to maintain survival when facing environmental stresses. As salt stress restricts crop architecture and grain yield, identifying genetic variations associated with growth and yield responses to salinity is critical for breeding optimal crop varieties. OsDSK2a is a pivotal modulator of plant growth and salt tolerance via the modulation of gibberellic acid (GA) metabolism; however, its regulation remains unclear. Here, we showed that OsDSK2a can be phosphorylated at the second amino acid (S2) to maintain its stability. The gene-edited mutant osdsk2aS2G showed decreased plant height and enhanced salt tolerance. SnRK1A modulated OsDSK2a-S2 phosphorylation and played a substantial role in GA metabolism. Genetic analysis indicated that SnRK1A functions upstream of OsDSK2a and affects plant growth and salt tolerance. Moreover, SnRK1A activity was suppressed under salt stress, resulting in decreased phosphorylation and abundance of OsDSK2a. Thus, SnRK1A preserves the stability of OsDSK2a to maintain plant growth under normal conditions, and reduces the abundance of OsDSK2a to limit growth under salt stress. Haplotype analysis using 3 K-RG data identified a natural variation in OsDSK2a-S2. The allele of OsDSK2a-G downregulates plant height and improves salt-inhibited grain yield. Thus, our findings revealed a new mechanism for OsDSK2a stability and provided a valuable target for crop breeding to overcome yield limitations under salinity stress.

Biofilm producing plant growth promoting bacteria in combination with glycine betaine uplift drought stress tolerance of maize plant.

The escalating threat of drought poses a significant challenge to sustainable food production and human health, as water scarcity adversely impacts various aspects of plant physiology. Maize, a cornerstone in staple cereal crops, faces the formidable challenge of drought stress that triggers a series of transformative responses in the plant. The present study was carried out in two sets of experiments. In first experiment, drought stress was applied after maintaining growth for 45 days and then irrigation was skipped, and plant samples were collected at 1st, 3rd and 6th day of drought interval for evaluation of changes in plant growth, water relation (relative water content) and antioxidants activity by inoculating indigenously isolated drought tolerant biofilm producing rhizobacterial isolates (Bacillus subtilis SRJ4, Curtobacterium citreum MJ1). In the second experiment, glycine betaine was applied as osmoregulator in addition to drought tolerant PGPR to perceive modulation in photosynthetic pigments (Chlorophyll a and b) and plant growth under varying moisture stress levels (100, 75 and 50% FC). Results of the study revealed upsurge in root and shoot length, fresh and dry biomass of root and shoot besides increasing chlorophyll contents in water stressed inoculated plants compared to uninoculated plants. Glycine betaine application resulted in an additional boost to plant growth and photosynthetic pigments, when applied in combination with bacterial inoculants. However, both bacterial inoculants behaved differently under drought stress as evident from their biochemical and physiological attributes. Isolate SRJ4 proved to be superior for its potential to express antioxidant activity, leaf water potential and relative water contents and drought responsive gene expression while isolate MJ1 showed exclusive increase in root dry biomass and plant P contents. Though it is quite difficult to isolate the bacterial isolates having both plant growth promoting traits and drought tolerance together yet, such biological resources could be an exceptional option to be applied for improving crop productivity and sustainable agriculture under abiotic stresses. By exploring the combined application of PGPR and glycine betaine, the study seeks to provide insights into potential strategies for developing sustainable agricultural practices aimed at improving crop resilience under challenging environmental conditions.

Determination, temporal variation and potential health risk assessment of pesticide residues in grapes from South and Southwest China

Pesticide residues in grapes from South and Southwest China were determined using the QuEChERS procedure and UHPLC-MS/MS and GC-MS/MS methods. The 4-year monitoring and survey showed 94.6% of the 1341 samples of grapes collected from eight main production areas contained one or multiple pesticide residues (above the respective LOQs). Overall, 40 pesticides were detected, including 24 fungicides, 12 insecticides, 2 acaricides and 2 plant growth modulators, of which one pesticide was unauthorised for use in treating grapes. Two or more pesticide residues were discovered in 87.4% of the samples (above the respective LOQs), and pesticide residues in 5.7% of the samples exceeded the MRLs, such as difenoconazole, cyhalothrin, propiconazole, etc. The main risk factors affecting the safety of grape before 2019 were difenoconazole, cyhalothrin and cyazofamid. After 2019, however, the frequency of occurrence of the above pesticides significantly declined, and the banned or restricted pesticides including omethoate were not found, which was credited to the stricter supervision and management policies by local governments. Despite the high detection rates and multi-residue occurrence of pesticides in grapes, about 84% of the samples were compliant with regulatory standards. Moreover, the accumulative chronic diet risk determined from ADI is very low. This study and timely monitoring can ensure that grape growers comply with GAP and minimise the occurrence of residues.

Growth modulation in Genu Valgum secondary to Multiple Hereditary Exostosis

Objective: Hereditary Multiple Exostoses (HME) is a rare disease with a prevalence of 1:50,000, which can manifest in various ways. This cartilaginous tumor can appear in a spectrum, ranging from mobility restriction to chronic pain and alignment disorders. Literature often describes forearm deformities such as excessive radial bowing, ulnar shortening, and radial head dislocation. Factors resulting in genu valgus and specific treatment strategies are not frequently studied. Considerations specific to the disease, such as the morphology of the lesion, time taken for deformity correction, and timing of exostosis excision, should be taken into account when treating genu valgum in patients with MHE. Case: Our case report highlights a teenager with underlying MHE who presented to us with "knock knees" since childhood. Clinically, it was observed that the patient had bilateral genu valgum and multiple bony swellings overlying the proximal tibia and distal femur. The deformity was subsequently addressed with bilateral proximal tibia hemi-epiphysiodesis, exostoses excision of bilateral proximal tibia, and left distal tibia. The radiographs and surgical technique are detailed in the case report. Our literature review indicates that distal femur exostoses play a significant role in causing genu valgus in these patients. The morphology of the exostoses also affects the development of genu valgus, with sessile lesions contributing more bony deformity compared to pedunculated lesions. Hemi-epiphysiodesis, as performed in our patient, often achieves outcomes similar to idiopathic genu valgum. However, the correction would take about 6 months longer, so in cases of MHE, correction should start earlier. Another dilemma in cases of pathological genu valgus is whether to remove exostosis during corrective surgery. Literature advocates delaying the excision of proximal tibia exostoses as long as possible to reduce the risk of accelerating deformity worsening. Conclusion: In conclusion, factors influencing genu valgus progression include the morphology of the lesion (sessile), presence of metaphyseal flaring, and a history of prior exostosis excision. Early detection of the pathology would yield better outcomes, as the rate of deformity correction in children with MHE is slower due to abnormal physis. Meticulous surgical technique to prevent injury to the physis and knee immobilization post-excision of exostosis are important factors in preventing the development of genu valgus in children with MHE.

Tirzepatide prevents neurodegeneration through multiple molecular pathways

BackgroundSeveral evidence demonstrated that glucagon-like peptide 1 receptor agonists (GLP1-RAs) reduce the risk of dementia in type 2 diabetes patients by improving memory, learning, and overcoming cognitive impairment. In this study, we elucidated the molecular processes underlying the protective effect of Tirzepatide (TIR), a dual glucose-dependent insulinotropic polypeptide receptor agonist (GIP-RA)/ GLP-1RA, against learning and memory disorders.MethodsWe investigated the effects of TIR on markers of neuronal growth (CREB and BDNF), apoptosis (BAX/Bcl2 ratio) differentiation (pAkt, MAP2, GAP43, and AGBL4), and insulin resistance (GLUT1, GLUT4, GLUT3 and SORBS1) in a neuroblastoma cell line (SHSY5Y) exposed to normal and high glucose concentration. The potential role on DNA methylation of genes involved in neuroprotection and epigenetic modulators of neuronal growth (miRNA 34a), apoptosis (miRNA 212), and differentiation (miRNA 29c) was also investigated. The cell proliferation was detected by measuring Ki-67 through flow cytometry. The data were analysed by SPSS IBM Version 23 or GraphPad Prism 7.0 software and expressed as the means ± SEM. Differences between the mean values were considered significant at a p-value of < 0.05. GraphPad Prism software was used for drawing figures.ResultsFor the first time, it was highlighted: (a) the role of TIR in the activation of the pAkt/CREB/BDNF pathway and the downstream signaling cascade; (b) TIR efficacy in neuroprotection; (c) TIR counteracting of hyperglycemia and insulin resistance-related effects at the neuronal level.ConclusionsWe demonstrated that TIR can ameliorate high glucose-induced neurodegeneration and overcome neuronal insulin resistance. Thus, this study provides new insight into the potential role of TIR in improving diabetes-related neuropathy.Graphical

Phylogenomic curation of Ovate Family Proteins (OFPs) in the U's Triangle of Brassica L. indicates stress-induced growth modulation.

The Ovate Family Proteins (OFPs) gene family houses a class of proteins that are involved in regulating plant growth and development. To date, there is no report of the simultaneous functional characterization of this gene family in all members of U's Triangle of Brassica. Here, we retrieved a combined total of 256 OFP protein sequences and analyzed their chromosomal localization, gene structure, conserved protein motif domains, and the pattern of cis-acting regulatory elements. The abundance of light-responsive elements like G-box, MRE, and GT1 motif suggests that OFPs are sensitive to the stimuli of light. The protein-protein interaction network analysis revealed that OFP05 and its orthologous genes were involved in regulating the process of transcriptional repression through their interaction with homeodomain transcription factors like KNAT and BLH. The presence of domains like DNA binding 2 and its superfamily speculated the involvement of OFPs in regulating gene expression. The biotic and abiotic stress, and the tissue-specific expression analysis of the RNA-seq datasets revealed that some of the genes such as BjuOFP30, and BnaOFP27, BolOFP11, and BolOFP10 were highly upregulated in seed coat at the mature stage and roots under various chemical stress conditions respectively which suggests their crucial role in plant growth and development processes. Experimental validation of prominent BnaOFPs such as BnaOFP27 confirmed their involvement in regulating gene expression under salinity, heavy metal, drought, heat, and cold stress. The GO and KEGG pathway enrichment analysis also sheds light on the involvement of OFPs in regulating plant growth and development. These findings have the potential to serve as a forerunner for future studies in terms of functionally diverse analysis of the OFP gene family in Brassica and other plant species.

Modulation in growth, oxidative stress, photosynthesis, and morphology reveals higher toxicity of alpha-cypermethrin than chlorpyrifos towards a non-target green alga at high doses

Considering the frequent detection of pesticides in the aquatic environment, the ecotoxicological effects of Chlorpyrifos (CHP), an organophosphate, and alpha-cypermethrin (ACM), a pyrethroid, on freshwater microalgae were compared for the first time in this study. High concentrations of both CHP and ACM significantly suppressed the growth of test microalga Graesiella emersonii (p < 0.05). The 96-h EC50 of CHP and ACM were 54.42 mg L−1 and 29.40 mg L−1, respectively. Sub-inhibitory doses of both pesticides increased ROS formation in a concentration-dependent manner, which was accompanied by changes in antioxidant enzymes activities, lipid peroxidation, and variations in photosynthetic pigment concentration. Furthermore, both pesticides influenced photosystem II performance, oxygen-evolving complex efficiency and, intracellular ATP levels. Scanning electron microscopy analysis revealed that high concentrations of both CHP and ACM caused considerable morphological changes in the microalga. In comparison, CHP was more toxic than ACM at low concentrations, whereas ACM was more toxic at high concentrations.

Recent insights into the role of hormones during development and their functional regulation.

Hormones play a vital role in development from conception to birth and throughout the human lifespan. These periods are logically divided into fetal development, pre-pubertal growth, puberty, and adulthood. Deviations from standard physiological levels and release patterns of constituent hormones can lead to pathology affecting the normal developmental trajectory. Research is ongoing to better understand the mechanisms of these hormones and how their modulation affects development. This article focuses on recent developments in understanding the role hormones play in development. We also cover recent discoveries in signaling pathways and hormonal regulation. New and continuing research into functional hormone regulation focuses on sex hormones, gonadotropic hormones, growth hormones, insulin-like growth factor, thyroid hormone, and the interconnectedness of each of these functional axes. Currently, the abundance of work focuses on fertility and correction of sex hormone levels based on an individual's condition and stage in life. Continuing research is needed to fully understand the long-term effects of hormone modulation in growth and sexual development. The role of each hormone in parallel endocrine axes should also be more thoroughly investigated to help improve the safety and efficacy in endocrine pharmacotherapeutics.

5β-reduced neuroactive steroids as modulators of growth and viability of postnatal neurons and glia

Endogenous neurosteroids (NS) and their synthetic analogs, neuroactive steroids (NAS), are potentially useful drug-like compounds affecting the pathophysiology of miscellaneous central nervous system disorders (e.g. Alzheimer´s disease, epilepsy, depression, etc.). Additionally, NS have been shown to promote neuron viability and neurite outgrowth upon injury. The molecular, structural and physicochemical basis of the NS effect on neurons is so far not fully understood, and the development of new, biologically relevant assays is essential for their comparative analysis and for assessment of their mechanism of action. Here, we report the development of a novel, plate-based, high-content in vitro assay for screening of NS and newly synthesized, 5β-reduced NAS for the promotion of postnatal neuron survival and neurite growth using fluorescent, postnatal mixed cortical neuron cultures isolated from thy1-YFP transgenic mice. The screen allows a detailed time course analysis of different parameters, such as the number of neurons or neurite lengths of 7-day, in vitro neuron cultures. Using the screen, we identify a new NAS, compound 42, that promotes the survival and growth of postnatal neurons significantly better than several endogenous NS (dehydroepiandrosterone, progesterone, and allopregnanolone). Interestingly, we demonstrate that compound 42 also promotes the proliferation of glia (in particular oligodendrocytes) and that the glial function is critical for its neuron growth support. Computational analysis of the biological data and calculated physicochemical properties of tested NS and NAS demonstrated that their biological activity is proportional to their lipophilicity. Together, the screen proves useful for the selection of neuron-active NAS and the comparative evaluation of their biologically relevant structural and physicochemical features.

Sydnone Imines as a New Class of Promising Plant Growth and Stress Tolerance Modulators—A First Experimental Structure–Activity Overview

Due to the oncoming climate changes, various environmental stresses (drought, salinity, heavy-metals, low or high temperatures, etc.) might dramatically affect crop yields and the quality of produced foods. Therefore, to meet the growing food demand of the human population, improvement of stress tolerance of the currently cultured crops is required. The knowledge of the molecular underlying mechanisms provides a versatile instrument to correct plant metabolism via chemical tools and to thereby increase their adaptive potential. This will preserve crop productivity and quality under abiotic stress conditions. Endogenously produced nitric oxide (NO) is one of the key signaling factors activating stress tolerance mechanisms in plants. Thus, the application of synthetic NO donors as stress-protective phytoeffectors might support maintaining plant growth and productivity under stressful conditions. Sydnone imines (sydnonimines) are a class of clinically established mesoionic heterocyclic NO donors which represent a promising candidate group for such phytoeffectors. Therefore, here, we provide an overview of the current progress in the application of sydnone imines as exogenous NO donors in plants, with a special emphasis on their potential as herbicides as well as herbicide antidotes, growth stimulants and stress protectors triggering plant tolerance mechanisms. We specifically address the structure–activity relationships in the context of the growth modulating activity of sydnone imines. Growth stimulating or antidote effects are typical for 4-α-hydroxybenzyl derivatives of sydnone imines containing an alkyl substituent in position N-3. The nature of the substituent of the N-6 atom has a significant influence on the activity profile and the intensity of the effect. Nevertheless, further investigations are necessary to establish reliable structure–activity relationships (SAR). Consequently, sydnone imines might be considered promising phytoeffector candidates, which are expected to exert either protective effects on plants growing under unfavorable conditions, or herbicidal ones, depending on the exact structure.

Efficacy of immune checkpoint inhibitors for metastatic colorectal cancer with microsatellite instability in second or latter line using synthetic control arms: A non-randomised evaluation

PurposeImmune checkpoint inhibitors (ICIs) appeared active in single-arm trials for patients with chemoresistant metastatic colorectal cancer (mCRC) harboring microsatellite instability (MSI). Given the paucity of randomised controlled trials (RCTs) in this setting, we evaluated the effect size of ICIs using intra-patients comparison and ARCAD database as historical controls. Patients and methodsIndividual-patient data from NIPICOL and CheckMate 142 phase II trials that evaluated a combination of ICIs for MSI mCRC patients (N = 176) and from five non-ICI mCRC historical RCTs in second-line or latter (N = 4026) were analyzed. Firstly, promising of ICIs was identified using intra-patient comparison based on growth modulation index (GMI) defined the ratio of progression-free survivals (PFS) on ICIs and previous line of therapy. Survival outcomes of ICIs-treated patients were then compared with those matched non-ICIs treated from ARCAD database historical RCTs. ResultsAmong ICIs-treated patients, median PFS on ICIs was 32.66 (range 0.10–74.25) versus 4.07 months (range 0.7–49.87) on prior therapy, resulting on median GMI of 4.97 (range 0.07–59.51; hazard-ratio (HR)= 0.16 (95 %CI=0.11–0.22, P < 0.001)). Compared to matched non-ICI patients, in third-line, median overall survival (OS) was not reached with ICIs versus 3.52 months with placebo (HR=0.20, 95 %CI=0.10–0.41, P < 0.001), and 6.51 months with active drugs (HR=0.30, 95 %CI=0.15–0.60, P = 0.001). In second-line, median OS was not reached with ICIs versus 11.7 months with chemotherapy+placebo (HR=0.12, 95 %CI=0.07–0.22, P < 0.001), and 16.3 months with chemotherapy+targeted therapy (HR=0.10, 95 %CI=0.05–0.19, P < 0.001). ConclusionICIs demonstrates high effect size for MSI mCRC patients in second-line and later. This work might be useful as an example of methodology to avoid RCTs when benefit from experimental therapy is likely to be high.

Growth and longevity modulation through larval environment mediate immunosenescence and immune strategy of Tenebrio molitor

BackgroundThe Disposable Soma Theory of aging suggests a trade-off between energy allocation for growth, reproduction and somatic maintenance, including immunity. While trade-offs between reproduction and immunity are well documented, those involving growth remain under-explored. Rapid growth might deplete resources, reducing investment in maintenance, potentially leading to earlier or faster senescence and a shorter lifespan. However, rapid growth could limit exposure to parasitism before reaching adulthood, decreasing immunity needs. The insect immunity’s components (cellular, enzymatic, and antibacterial) vary in cost, effectiveness, and duration. Despite overall immunity decline (immunosenescence), its components seem to age differently. We hypothesize that investment in these immune components is adjusted based on the resource cost of growth, longevity, and the associated risk of parasitism.ResultsWe tested this hypothesis using the mealworm beetle, Tenebrio molitor as our experimental subject. By manipulating the larval environment, including three different temperatures and three relative humidity levels, we achieved a wide range of growth durations and longevities. Our main focus was on the relationship between growth duration, longevity, and specific immune components: hemocyte count, phenoloxidase activity, and antibacterial activity. We measured these immune parameters both before and after exposing the individuals to a standard bacterial immune challenge, enabling us to assess immune responses. These measurements were taken in both young and older adult beetles. Upon altering growth duration and longevity by modifying larval temperature, we observed a more pronounced investment in cellular and antibacterial defenses among individuals with slow growth and extended lifespans. Intriguingly, slower-growing and long-lived beetles exhibited reduced enzymatic activity. Similar results were found when manipulating larval growth duration and adult longevity through variations in relative humidity, with a particular focus on antibacterial activity.ConclusionThe impact of growth manipulation on immune senescence varies by the specific immune parameter under consideration. Yet, in slow-growing T. molitor, a clear decline in cellular and antibacterial immune responses with age was observed. This decline can be linked to their initially stronger immune response in early life. Furthermore, our study suggests an immune strategy favoring enhanced antibacterial activity among slow-growing and long-lived T. molitor individuals.

Investigation of stimulated growth effect by application of L-aspartic acid on poplar

Poplar is an important resource for the production of wood-based products. L-aspartic acid (Asp) is a proteinogenesis amino acid that plays a critical role in cell growth modulation and stress response. Also, Asp is widespread in the commonly used amino acid-based biostimulant products. However, its effects on poplars remain largely unknown. To address this issue, we applied different concentrations of Asp to poplar seedlings under N reduction conditions (simulating low N availability status), and assessed the effects of Asp on poplar growth and development. Our results showed that supplement of Asp at an appropriate concentration, that is, 0.5 mM in the present study, had the best growth-promoting effect on poplars. It significantly increased poplar biomass and height, and simultaneously elevated the photosynthetic parameters, the chlorophyll content and total free amino acid content, as well as the activity of N metabolism-related enzymes in poplars-treated with Asp in comparision with the control plants without Asp addition. Furthermore, the expression of Asp-metabolism pathway genes (AspAT1, AspAT6, AspAT10, AK1, ASS1 and ASS2), key chlorophyll synthesis gene (LHCB3) and cell division gene (CYCD3) was all dramatically upregulated in response to Asp application, suggesting that Asp-induced effect invoked photosynthesis activation and metabolism reprogramming. More interestingly, our results showed that the levels of various phytohormones, such as auxin and cytokinins were remarkably altered, and key auxin transport gene (PIN1c-2), auxin conjugation gene (GH3.6–2) were induced by Asp treatment, indicative of the possible involvement of plant hormones under the effect of Asp, especially fine-tuning auxin homeostasis by forming IAA-Asp conjugates to store auxin and catabolizing IAA-Asp to release auxin, may account for Asp-mediated growth modulation to an extent in poplars. Our study provides a novo insight into the growth promotion effect of Asp on poplars, especially, when the N source is limited, even though the underlying mechanism of Asp-induced stimulation action needs further in-depth investigation.