Stimulatory Effect Research Articles

Integrative analysis of transcriptome and phytohormones reveals new genes network involved in shoot apical meristem development in maize under blue and red light

Blue and red light play a significant role in regulating transcriptional expression, phytohormone levels, and gene signal network reconstruction in the shoot apical meristem. To further elucidate the effects of red and blue light on transcriptional characteristics and phytohormone expression, and to explore signaling pathways and novel genes, we conducted comparative experiments under different light quality treatments. The results are as follows: a total of 22,327 genes were found to be effectively expressed, with a considerable number of alternative splicing events occurring concurrently. Furthermore, 1063 differentially expressed genes were identified, primarily associated with transcriptional variations resulting from the transition from blue light to red light. These differentially expressed genes are predominantly involved in KEGG annotations related to photosynthesis, DNA replication and carbon metabolism. The findings from targeted metabolism analysis reveal significant variations in 45 phytohormones, including ABA, auxin, CK, ETH, GA, JA, and SA. Correlation analysis between transcription and metabolism further indicates that IAA.Glc, IAA.Glu, TRP, mT9G, and GA53 show the strongest correlation with differentially expressed genes, with genes ssu1, ssu2, pep1, me3, and hsp90 displaying a significant relationship. Additionally, in the protein–protein interaction network, three novel genes (LOC100286132, LOC100510780, and LOC103653934) were identified as significant responding to the regulation of blue and red light. Following protein function analysis, the proteins were provisionally identified as ubiquitin conjugating enzyme-like, OXIDATIVE STRESS 3LIKE 2-like, and Ribosomal L 18p/L5e, respectively. Conclusively, it is depicted that the most significant impact of transitions between red light, blue light, and two light qualities lies in the transcriptional process of shot apical meristem of the C4 plant maize. This process is intricately linked to nitrogen availability, carbon dioxide assimilation capacity, and the stimulatory effects of auxin, cytokinin, and gibberellin. The expression of the putative L 18/L5e in blue and BTR light conditions may enhance transcript and synthesis capabilities of ribosome.

Effect of Fertilizers, Seeding Rates of the Legume Component and Weather Conditions on Yield and Accumulation of Plant Residues of LupineCereal Mixtures on Grain in the Center of the NonChernozem Region

On mediumcultivated mediumloamy sod-podzolic soils of the Central NonChernozem region, well provided with mobile phosphorus and potassium (IV–V class), the cultivation of lupine–cereal mixtures for grain with the participation of narrowleaved lupine varieties Ladny, spring wheat, barley and oats varieties of Nemchin breeding after grain precursors in a changing climate ensured grain production in an average of 5 up to 3.0–3.7 t/ha with a share of the legume component of 29–57% and the remaining dry mass of plant residues in the soil up to 6–8 t/ha with the accumulation of total nitrogen in it up to 40–60 kg/ha and biological – 12–17 kg/ha. In arid conditions (GTK = 0.80–0.92), a mixture of lupine and barley was distinguished for the better in terms of yield and weight of plant residues, and in case of waterlogging (GTK = 2.47) – mixtures with oats and spring wheat. The inclusion of liquid micronutrients of organic nature in the technological process with a stimulating effect for non-root top dressing eliminated the need for pre-sowing application of nitrogen fertilizer and, in conditions of moderate aridity, increased grain yield, depending on the composition of the mixture, to 3.9–4.9 t/ha (by 53–59%), the yield of plant residues – up to 7–11 t/ha (by 54–139%) and in them there is an accumulation of symbiotically bound nitrogen – up to 15–25 kg/ha or 5–8 times more than without their use.

Zinc Oxide Nanoparticles in the “Soil–Bacterial Community–Plant” System: Impact on the Stability of Soil Ecosystems

The use of man-made nanoparticles (NPs) has increased exponentially in recent years, many of which accumulate in significant quantities in soil, including through use in agriculture as nanofertilizers and nanopesticides. ZnO NPs are more environmentally friendly but have specific antimicrobial activity, which can affect soil microbiota, thereby influencing key microbial processes such as mineralization, nitrogen fixation and plant growth-promoting activities. Their behavior and persistence in soil depend on their chemical nature and soil characteristics. This review summarizes the applications of ZnO NPs in soil systems and their effects on various plants and soil microorganisms, particularly rhizobacteria that promote plant growth. A stimulating effect of ZnO NPs on the morphometric and biochemical characteristics of plants, as well as on soil microbiota and its activity at relatively low concentrations of up to 500 mg/mL and 250 mg/kg, respectively, is observed. As the concentration of ZnO NPs increases above these limits, toxic effects appear. The different effects of ZnO NPs are related to their size, dose, duration of exposure, solubility in water, as well as soil type, acidity and organic matter content. The review substantiates the need to study the behavior of ZnO NPs in the “soil-plant-microbiota” system for the possibility of using nanotechnologies in the agricultural industry and ensuring the safety of agricultural products.

Development of functional probiotic yogurt from buffalo milk supplemented with red beetroot (Beta vulgaris L.) as an antioxidant, natural colorant, and starter growth stimulant

The physicochemical, sensorial, and probiotics viability properties of supplemented probiotic yogurt with beet juice (BJ) at various ratios (0–5)% in fresh and stored yogurts were determined. 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity (%) and minerals content of BJ-yogurts were increased. The color aspects (ΔE) were improved with the increasing of BJ. BJ-yogurts at 1 and 3 % were color-stable when stored in the refrigerator (5 ± 1 °C) for 9 days. The syneresis reached of 42.9, 43.0, 43.5, and 43.5 mL for control, 1 %, 3 %, and 5 % BJ, respectively in stored yogurts for 12 days. Moreover, the probiotic yogurt viability in BJ-yogurts treatments was found to be higher (p < 0.05) than control which recorded of 7.0 ± 0.08, 7.22±0.04, 7.34±0.002 log cfu/mL for streptococci, lactobacilli, and bifidobacteria counts respectively in 5 % BJ treatment, on the other hand, in control recorded of 6.76±0.07, 7.11±0.06, 6.66±0.030 log cfu/mL for streptococci, lactobacilli, and bifidobacteria counts respectively; indicating that BJ had a stimulating effect on the additive starter. Fresh yogurts with 1 % BJ had superior sensorial scores than other treatments, and generally maintained desirable organoleptic attributes for up to 9 days. This study provides a nutritious, low-cost, stable, and attractive colored yogurts from buffalo milk without negative impact on physiochemical qualities.

MiR-155 deficiency and hypoxia results in metabolism switch in the leukemic B-cells

Hypoxia represents one of the key factors that stimulates the growth of leukemic cells in their niche. Leukemic cells in hypoxic conditions are forced to reprogram their original transcriptome, miRNome, and metabolome. How the coupling of microRNAs (miRNAs)/mRNAs helps to maintain or progress the leukemic status is still not fully described. MiRNAs regulate practically all biological processes within cells and play a crucial role in the development/progression of leukemia. In the present study, we aimed to uncover the impact of hsa-miR-155-5p (miR-155, MIR155HG) on the metabolism, proliferation, and mRNA/miRNA network of human chronic lymphocytic leukemia cells (CLL) in hypoxic conditions. As a model of CLL, we used the human MEC-1 cell line where we deleted mature miR-155 with CRISPR/Cas9. We determined that miR-155 deficiency in leukemic MEC-1 cells results in lower proliferation even in hypoxic conditions in comparison to MEC-1 control cells. Additionally, in MEC-1 miR-155 deficient cells we observed decreased number of populations of cells in S phase. The miR-155 deficiency under hypoxic conditions was accompanied by an increased apoptosis. We detected a stimulatory effect of miR-155 deficiency and hypoxia at the level of gene expression, seen in significant overexpression of EGLN1, GLUT1, GLUT3 in MEC-1 miR-155 deficient cells. MiR-155 deficiency and hypoxia resulted in increase of glucose and lactate uptake. Pyruvate, ETC and ATP were reduced. To conclude, miR-155 deficiency and hypoxia affects glucose and lactate metabolism by stimulating the expression of glucose transporters as GLUT1, GLUT3, and EGLN1 [Hypoxia-inducible factor prolyl hydroxylase 2 (HIF-PH2)] genes in the MEC-1 cells.

Ropivacaine synergizes with sorafenib to induce apoptosis of hepatocellular carcinoma cells via the IL-6/STAT3 pathway.

The development of resistance in hepatocellular carcinoma (HCC) cells limits the effectiveness of sorafenib, but combination therapy with other drugs may have a positive effect. However, the effect of ropivacaine combined with sorafenib on the treatment of HCC cells and its potential regulatory mechanisms remain unclear. The proliferation and apoptosis of HCC cells treated with ropivacaine, sorafenib, and ropivacaine plus sorafenib were analyzed by cell-counting kit 8 and flow cytometry. The protein levels were measured by Western blot. The antitumor effect of ropivacaine, sorafenib, and their combination was verified by a tumor xenograft model. Ropivacaine and sorafenib markedly impeded the viability of HCC cells in a concentration-dependent manner. Compared with ropivacaine or sorafenib treatment alone, ropivacaine and sorafenib combination treatment impeded HCC cell proliferation, facilitated apoptosis, enhanced cleaved caspase-3, cleaved caspase-9, and cyclin D1 protein expression, while it reduced IL-6 and p-STAT3 expression and inhibited tumor growth invivo. Importantly, the activation of the IL-6/STAT3 pathway could reverse the repressive or stimulative effects of ropivacaine and sorafenib on the proliferation and apoptosis in HCC cells. In summary, ropivacaine synergistically induces sorafenib-stimulated apoptosis of HCC cells via the IL-6/STAT3 pathway. Ropivacaine is a potential drug for the treatment of HCC when combined with sorafenib.

Inter-individual variability in neural response to low doses of LSD

The repeated use of small doses of psychedelics (also referred to as “microdosing”) to facilitate benefits in mental health, cognition, and mood is a trending practice. Placebo-controlled studies however have largely failed to demonstrate strong benefits, possibly because of large inter-individual response variability. The current study tested the hypothesis that effects of low doses of LSD on arousal, attention and memory depend on an individual’s cognitive state at baseline. Healthy participants (N = 53) were randomly assigned to receive repeated doses of LSD (15 mcg) or placebo on 4 occasions divided over 2 weeks. Each treatment condition also consisted of a baseline and a 1-week follow-up visit. Neurophysiological measures of arousal (resting state EEG), pre-attentive processing (auditory oddball task), and perceptual learning and memory (visual long-term potentiation (LTP) paradigm) were assessed at baseline, dosing session 1 and 4, and follow-up. LSD produced stimulatory effects as reflected by a reduction in resting state EEG delta, theta, and alpha power, and enhanced pre-attentive processing during the acute dosing sessions. LSD also blunted the induction of LTP on dosing session 4. Stimulatory effects of LSD were strongest in individuals with low arousal and attention at baseline, while inhibitory effects were strongest in high memory performers at baseline. Decrements in delta EEG power and enhanced pre-attentive processing in the LSD treatment condition were still present during the 1-week follow-up. The current study demonstrates across three cognitive domains, that acute responses to low doses of LSD depend on the baseline state and provides some support for LSD induced neuroadaptations that sustain beyond treatment.

Ionizable STING-Activating Nanoadjuvants Enhance Tumor Immunogenicity and Potentiate Immunotherapy Efficacy in Solid Tumors.

Therapeutic strategies that induce inflammatory responses in immunologically "cold" tumors have the potential to improve immunotherapeutic outcomes. Pharmacologically activating the STING pathway induces innate immunity, subsequently enhancing tumor immunogenicity. Here, we developed a nanoadjuvant with tumor-restricted pharmacology that rapidly activated STING and reshaped the tumor microenvironment (TME). The non-nucleotide STING agonist MSA-2 was chemically engineered with a piperazine motif linked by a saturated hydrocarbon chain of varying lengths to produce ionizable prodrugs that were further developed into nanoadjuvants. Compared with state-of-the-art liposomes, the nanoadjuvant displayed prolonged retention in the circulation and improved intratumoral delivery. In the acidic TME, the nanoadjuvant underwent polyethylene glycol deshielding, enabling efficient extravasation and penetration into tumors. Concomitantly, the STING prodrug escaped from the endo/lysosome compartment to partition into the cytosol for spontaneous esterase-catalyzed drug activation. In mouse models of syngeneic and chemically induced colorectal cancers, nanoparticle treatment provoked robust STING-mediated antitumor immunity, shifting the tumor immune landscape from immunosuppressed to tumoricidal. Additionally, the nanoadjuvant demonstrated antitumor efficacy in triple-negative breast cancer, which was further enhanced by the addition of immune checkpoint inhibitors. Collectively, this study demonstrates the safety and immune stimulating effects of a STING-activating nanoadjuvant, supporting the clinical evaluation of this STING immunotherapeutic alone and in combination with other immunotherapies.

Amino acids biosynthesis in root hair development: a mini-review.

Metabolic factors are essential for developmental biology of an organism. In plants, roots fulfill important functions, in part due to the development of specific epidermal cells, called hair cells that form root hairs (RHs) responsible for water and mineral uptake. RH development consists in (a) patterning processes involved in formation of hair and non-hair cells developed from trichoblasts and atrichoblasts; (b) RH initiation; and (c) apical (tip) growth of the RH. Here we review how these processes depend on pools of different amino acids and what is known about RH phenotypes of mutants disrupted in amino acid biosynthesis. This analysis shows that some amino acids, particularly aromatic ones, are required for RH apical (tip) growth, and that not much is known about the role of amino acids at earlier stages of RH formation. We also address the role of amino acids in rhizosphere, inhibitory and stimulating effects of amino acids on RH growth, amino acids as N source in plant nutrition, and amino acid transporters and their expression in the RHs. Amino acids form conjugates with auxin, a hormone essential for RH growth, and respective genes are overviewed. Finally, we outline missing links and envision some perspectives in the field.

Effects of copper, and aluminium in ionic, and nanoparticulate form on growth rate and gene expression of Setaria italica seedlings

This study aims to determine the effects of copper, copper oxide nanoparticles, aluminium, and aluminium oxide nanoparticles on the growth rate and expression of ACT-1, CDPK, LIP, NFC, P5CR, P5CS, GR, and SiZIP1 genes in five days old seedling of Setaria italica ssp. maxima, cultivated in hydroponic culture. Depending on their concentration (ranging from 0.1 to 1.8 mg L−1), all tested substances had both stimulating and inhibiting effects on the growth rate of the seedlings. Copper and copper oxide-NPs had generally a stimulating effect whereas aluminium and aluminium oxide-NPs at first had a positive effect but in higher concentrations they inhibited the growth. Treating the seedlings with 0.4 mg L−1 of each tested toxicant was mostly stimulating to the expression of the genes and reduced the differences between the transcript levels of the coleoptiles and roots. Increasing concentrations of the tested substances had both stimulating and inhibiting effects on the expression levels of the genes. The highest expression levels were usually noted at concentrations between 0.4 and 1.0 mg/L of each metal and metal nanoparticle, except for SiZIP1, which had the highest transcript amount at 1.6 mg L−1 of Cu2+ and at 0.1–0.8 mg L−1 of CuO-NPs, and LIP and GR from the seedling treated with Al2O3-NPs at concentrations of 0.1 and 1.6 mg L−1, respectively.

Effects of copper, and aluminium in ionic, and nanoparticulate form on growth rate and gene expression of Setaria italica seedlings

This study aims to determine the effects of copper, copper oxide nanoparticles, aluminium, and aluminium oxide nanoparticles on the growth rate and expression of ACT-1, CDPK, LIP, NFC, P5CR, P5CS, GR, and SiZIP1 genes in five days old seedling of Setaria italica ssp. maxima, cultivated in hydroponic culture. Depending on their concentration (ranging from 0.1 to 1.8 mg L−1), all tested substances had both stimulating and inhibiting effects on the growth rate of the seedlings. Copper and copper oxide-NPs had generally a stimulating effect whereas aluminium and aluminium oxide-NPs at first had a positive effect but in higher concentrations they inhibited the growth. Treating the seedlings with 0.4 mg L−1 of each tested toxicant was mostly stimulating to the expression of the genes and reduced the differences between the transcript levels of the coleoptiles and roots. Increasing concentrations of the tested substances had both stimulating and inhibiting effects on the expression levels of the genes. The highest expression levels were usually noted at concentrations between 0.4 and 1.0 mg/L of each metal and metal nanoparticle, except for SiZIP1, which had the highest transcript amount at 1.6 mg L−1 of Cu2+ and at 0.1–0.8 mg L−1 of CuO-NPs, and LIP and GR from the seedling treated with Al2O3-NPs at concentrations of 0.1 and 1.6 mg L−1, respectively.

Exosomal lncRNA HCP5 derived from human bone marrow mesenchymal stem cells improves chronic periodontitis by miR-24-3p/HO1/P38/ELK1 pathway

ObjectiveThe present study aimed to explore the function of human bone marrow mesenchymal stem cells (hBMMSCs)-derived exosomal long noncoding RNA histocompatibility leukocyte antigen complex P5 (HCP5) in the osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) to improve chronic periodontitis (CP). MethodsExosomes were extracted from hBMMSCs. Alizarin red S staining was used to detect mineralised nodules. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to measure HCP5 and miR-24-3p expression. The mRNA and protein levels of alkaline phosphatase (ALP), osteocalcin, osterix, runt-related transcription factor 2, bone morphogenetic protein 2, osteopontin, fibronectin, collagen 1, heme oxygenase 1 (HO1), P38, and ETS transcription factor ELK1 (ELK1) were detected using RT-qPCR and Western blot. Enzyme-linked immunosorbent assay (ELISA) kits were used to determine the HO1 and carbon monoxide concentrations. Heme, biliverdin, and Fe2+ levels were determined using detection kits. Micro-computed tomography, hematoxylin and eosin staining, ALP staining, tartrate-resistant acid phosphatase staining, ELISA, and RT-qPCR were conducted to evaluate the effect of HCP5 on CP mice. Dual luciferase, RNA immunoprecipitation, and RNA pulldown experiments were performed to identify the interactions among HCP5, miR-24-3p, and HO1. ResultsThe osteogenic ability of hPDLSCs significantly increased when co-cultured with hBMMSCs or hBMMSCs exosomes. Overexpression of HCP5 and HO1 in hBMMSCs exosomes promoted the osteogenic differentiation of hPDLSCs, and knockdown of HCP5 repressed the osteogenic differentiation of hPDLSCs. HCP5 knockdown enhanced the inflammatory response and repressed osteogenesis in CP mice. MiR-24-3p overexpression diminished the stimulatory effect of HCP5 on the osteogenic ability of hPDLSCs. Mechanistically, HCP5 acted as a sponge for miR-24-3p and regulated HO1 expression, and HO1 activated the P38/ELK1 pathway. ConclusionHBMMSCs-derived exosomal HCP5 promotes the osteogenic differentiation of hPDLSCs and alleviates CP by regulating the miR-24-3p/HO1/P38/ELK1 signalling pathway.

Intermittent Fasting Targets Osteocyte Neuropeptide Y to Relieve Osteoarthritis.

Osteoarthritis is a highly prevalent progressive joint disease that still requires an optimal therapeutic approach. Intermittent fasting is an attractive dieting strategy for improving health. Here this study shows that intermittent fasting potently relieves medial meniscus (DMM)- or natural aging-induced osteoarthritic phenotypes. Osteocytes, the most abundant bone cells, secrete excess neuropeptide Y (NPY) during osteoarthritis, and this alteration can be altered by intermittent fasting. Both NPY and the NPY-abundant culture medium of osteocytes (OCY-CM) from osteoarthritic mice possess pro-inflammatory, pro-osteoclastic, and pro-neurite outgrowth effects, while OCY-CM from the intermittent fasting-treated osteoarthritic mice fails to induce significant stimulatory effects on inflammation, osteoclast formation, and neurite outgrowth. Depletion of osteocyte NPY significantly attenuates DMM-induced osteoarthritis and abolishes the benefits of intermittent fasting on osteoarthritis. This study suggests that osteocyte NPY is a key contributing factor in the pathogenesis of osteoarthritis and intermittent fasting represents a promising nonpharmacological antiosteoarthritis method by targeting osteocyte NPY.

EFFECTS OF BIOLOGICALS AND PLANT GROWTH REGULATORS ON THE SOWING QUALITY OF EGGPLANT SEEDS

Purpose. To investigate effects of biologicals with fungicidal and stimulating activities and natural and synthetic plant growth regulators on the sowing quality of seeds of various eggplant hybrids. Methods. Laboratory methods in compliance with DSTU 4138:2002. Results. Soaking of seeds of various eggplant hybrids in Phytocid, Mycosan-B, PhytoHelp, MycoHelp, Azotophyt, Vympel, Humisol, Ivin, and Emistim C solutions had positive effects on the sowing quality of seed: the germination energy increased on average by 1–34% and the laboratory germinability – by 2–28%. Regardless of the hybrid, soaking of seeds in Phytocid, PhytoHelp, and MycoHelp solutions significantly increased the germination energy by 34.2%, 32.5%, and 32.5%, respectively, compared to the control and the laboratory germinability of seeds by 27.6%, 27, 4% and 27.4%, respectively. When plant growth regulators Azotophyt, Ivin, and Emistim C were applied, the germination energy and laboratory germinability increased on average by 18–28% and 15–24%, respectively, compared to the control. With Mycosan-B, Vympel, and Humisol, the germinability and germination energy of seeds did not differ from the control levels. Soaking of eggplant seeds in solutions of the studied formulations positively affected the seedling length: there was on average a 1.24- to 1.94-fold elongation of seedlings. It was found that seeds of the investigated eggplant hybrids were contaminated with phytopathogenic micromycetes and, depending on damage degree, the hybrids were ranked in the following order: 'Sapfir'&gt; 'Sharapova', 'Samurai' &gt; 'Leire' &gt; 'Night Lady', 'Fabina' &gt; 'Destan'. Mycosan-B, Vympel, and Humisol were little effective (2.5–6.3%) against phytopathogens in comparison with the other formulations. Phytocid, PhytoHelp, and MycoHelp exerted potent fungicidal effects, which varied on different hybrids: 42.5–68.8%, 30.0–65.6%, and 32.5–65.6%, respectively. The fungicidal effectiveness of Ivin and Emistim C was 20.3% and 18.8%, respectively. Conclusions. Soaking of eggplant seeds in solutions of biologicals (Phytocid, PhytoHelp, and MycoHelp) and synthetic and natural plant growth regulators (Ivin and Emistim C, respectively) was shown to beneficially affect the sowing quality of seeds and suppress fungal phytopathogens. The greatest stimulatory effects of Phytocid, PhytoHelp, MycoHelp, Ivin, and Emistim C on the sowing quality of seeds were seen on hybs. 'Destan', 'Leire', and 'Fabina'. 51–58% suppression of phytopathogens was achieved when eggplant seeds were soaked in Phytocid, PhytoHelp, and MycoHelp; Ivin and Emistim C resulted in 19–20% suppression. The greatest suppression of phytopathogens on eggplant seeds was observed on hybs. 'Destan', 'Leire', 'Night Lady', and 'Fabina'.

Agronomic and biochemical aspects of moringa dried leaf extract mediated growth and yield improvements in soybean

ABSTRACT Stimulatory effects of foliar application of moringa dried leaf extract (MDLE) at different growth stages were evaluated on soybean growth, physiology and productivity in a bioassay. The experiment comprised of two factors; foliar application of MDLE [T1: Water spray, T2: 0.5% w/v, T3: 1.0% and T4: 1.5%] and growth stages for foliar application, viz. S1: Vegetative stage (V4, four leaf stage) and S2: Reproductive stage (R1: onset of flowering). Foliar application of MDLE increased the growth, physiological, biochemical, yield and quality attributes of soybean. Notably, the application of 1.5% MDLE at the reproductive stage led to impressive boosts in seedling weight, chlorophyll contents, plant height, pod formation and seed yield. This concentration demonstrated the highest surge in gas exchange attributes and quality components like seed oil and protein contents. Significant increase in growth attributes (10–22%), photosynthetic pigments (7–10%), gas exchange attributes (21%), quality attributes (5–8%) and seed yield (27%) was recorded with 1.5% MDLE application. It is ascertained that the application of 1.5% MDLE at the reproductive stage improves soybean growth, yield and quality. This research provides quantitative confirmation of MDLE foliar application, emphasising its potential role as a natural growth enhancer for soybean crop.

Agronomic and biochemical aspects of moringa dried leaf extract mediated growth and yield improvements in soybean

ABSTRACT Stimulatory effects of foliar application of moringa dried leaf extract (MDLE) at different growth stages were evaluated on soybean growth, physiology and productivity in a bioassay. The experiment comprised of two factors; foliar application of MDLE [T1: Water spray, T2: 0.5% w/v, T3: 1.0% and T4: 1.5%] and growth stages for foliar application, viz. S1: Vegetative stage (V4, four leaf stage) and S2: Reproductive stage (R1: onset of flowering). Foliar application of MDLE increased the growth, physiological, biochemical, yield and quality attributes of soybean. Notably, the application of 1.5% MDLE at the reproductive stage led to impressive boosts in seedling weight, chlorophyll contents, plant height, pod formation and seed yield. This concentration demonstrated the highest surge in gas exchange attributes and quality components like seed oil and protein contents. Significant increase in growth attributes (10–22%), photosynthetic pigments (7–10%), gas exchange attributes (21%), quality attributes (5–8%) and seed yield (27%) was recorded with 1.5% MDLE application. It is ascertained that the application of 1.5% MDLE at the reproductive stage improves soybean growth, yield and quality. This research provides quantitative confirmation of MDLE foliar application, emphasising its potential role as a natural growth enhancer for soybean crop.

Black bioceramic scaffolds with micro/nano surface structure inducing mild hyperthermal environment for regenerating osteochondral defects

The treatment of osteochondral defects remains a huge challenge in clinic. The thermotherapy in traditional Chinese medicine is widely used in treating joint disease because the mild heat stimulation can promote the nutrient circulation. Inspired by the function of thermotherapy, we design a new thermotherapy strategy by using black bioceramic scaffolds to build local mild hyperthermal environment for treating osteochondral defects. Herein, black akermanite (B-AKT) bioceramic scaffolds with micro/nano surface structure and excellent photothermal conversion property are successfully fabricated. The temperature of scaffolds can be precisely controlled by near infrared (NIR) irradiation to construct a mild hyperthermal environment (～41°C). The micro/nanostructure on the scaffolds surface well promoted the cell adhesion and nutrition absorption by activating integrins and focal adhesion kinase (FAK). Meanwhile, the mild hyperthermal environment constructed by B-AKT scaffolds can simultaneously induce the chondrogenesis and osteogenesis both in vitro and in vivo by activating the expression of cellular heat shock proteins (HSPs) and corresponding signaling pathways. Furthermore, in rabbit osteochondral defects, mild hyperthermal environment induced by B-AKT scaffolds exhibits stimulatory effects on the integrated regeneration of cartilage and bone. This study proposes a novel therapy strategy of constructing local mild hyperthermal environment by black bioceramic scaffolds to treat osteochondral defects and other joint diseases.

P-336 Efficacy of Gonadotrophin (GnRH) antagonists for endometriosis-associated pelvic pain - A systematic review and meta-analysis

Abstract Study question Are Gonadotropin-releasing hormone (GnRH) antagonists effective and safe in managing pain associated with endometriosis? Summary answer GnRH antagonists seem effective and safe in managing endometriosis-associated pain for at least 12 weeks of use. There is uncertainty with the evidence. What is known already GnRH agonists are commonly used as effective second-line treatment for endometriosis-associated pain. However, these agonists have drawbacks and are not well-tolerated by many women. GnRH antagonists, offer potential advantages over the agonists while providing similar efficacy. They lack the initial stimulatory effects thereby avoiding the initial flare-up of symptoms, which is common with agonists. Furthermore, unlike agonists, which are mainly available as injections, the antagonist are also available as oral preparations, which allow titration of doses. In-vitro and animal studies have shown antagonists to induce regression of endometriotic lesions by apoptosis. However, their efficacy in relieving endometriosis-related pains remains uncertain. Study design, size, duration Extensive electronic search was conducted by the Cochrane Gynaecology and Fertility group, up to October 2023 to identify randomised controlled trials (RCTs) evaluating efficacy of GnRH antagonists in managing endometriosis-related pains compared with placebo and/or GnRH agonists. The main searched databases included CENTRAL, MEDLINE and EMBASE. Screening and selection of suitable studies were undertaken. Study eligibility criteria included treatment for ≥12 weeks, laparoscopically confirmed endometriosis and assessment of pelvic pain scores as the primary endpoint. Participants/materials, setting, methods Primary end-point was a change in endometriosis-related pain score after 12-week treatment with GnRH antagonists compared with placebo and/or GnRH agonists. Changes in pain and quality of life (QoL) scores and adverse-events of GnRH-antagonists were compared between the antagonist versus the agonist/placebo groups. Data were analysed using RevMan software (V5.4) to calculate the standard mean difference (SMD) for continuous data, and the risk ratios (RRs) with 95% confident intervals (95% CI) for categorical data. Main results and the role of chance Seven eligible RCTs (n = 3043) were included, all of which compared GnRH-antagonist versus placebo, and only three also compared versus GnRH-agonist (leuprorelin). The GnRH-antagonists included ASP1707, Elagolix, Linzagolix and Relugolix. Meta-analysis of overall pelvic pain data from seven RCTs (n = 1684) showed significantly greater reduction in pain scores after 12 weeks of GnRH-antagonist versus placebo at low (SMD -0.29 95%CI -0.39 to -0.19; I2=0) and high doses (SMD-0.53 95%CI -0.63 to -0.43, I2=38%). Similarly, meta-analysis of other pains (dysmenorrhoea, non-menstrual pain, and dyspareunia) showed significantly greater pain score reduction with GnRH-antagonists versus placebo. Meta-analysis of overall pelvic pain from two RCTs (n = 263) comparing GnRH-antagonist versus leuprorelin showed comparable reduction in pain scores after 12 weeks at low (SMD 0.33 95%CI 0.09 to 0.58, I2=44%) and high doses (SMD 0.06 95%CI -0.19 to 0.30, I2=0%). Similarly, meta-analysis of other forms of pain revealed comparable reductions of pain scores between both drugs. Meta-analysis of QoL data (2RCTs, n = 259) revealed significantly greater improvements in scores with leuprorelin versus antagonists at low (SMD 0.74 95% CI 0.46 to 1.01; I2=98%) and high doses (SMD 0.35, 95%CI, 0.09 to 0.60, I2=97%). The most frequently reported treatment-related side-effects were hot-flushes, headaches and nasopharyngitis. No life-threatening side-effects were reported. Limitations, reasons for caution There was high heterogeneity between studies (e.g. differences in antagonists and pain scoring systems). No information was provided on prior endometriosis surgery, which is a confounding factor. Additionally, declared COI by authors of all trials could introduce high risk of bias. Evidence certainty was therefore deemed low to very low. Wider implications of the findings GnRH-antagonists seem to offer a promising treatment for endometriosis-related pain, with comparable efficacy to the agonists, but with several potential advantages. However, given the low quality and uncertainty of the evidence in this meta-analysis, high quality RCTs with robust designs are needed before making any firm recommendations for clinical practice. Trial registration number not applicable

P-628 The role of Anti-Müllerian Hormone (AMH) on human osteoblast differentiation

Abstract Study question May AMH, an ovarian hormone belonging to the Transforming Growth Factor β superfamily (TGF-β), represent a possible candidate for use as a bone anabolic factor? Summary answer Our findings provided interesting clues on the stimulatory effects of AMH on mature osteoblasts expressing its specific receptor, the Anti-Müllerian Hormone Type-2 receptor (AMHRII). What is known already Impaired bone formation is a principal pathogenetic mechanism mediating bone fragility in osteoporosis. Several growth factors capable of inducing bone matrix apposition by increasing the number of osteoblast precursor cells or promoting the function of mature osteoblasts have been identified, including members of the TGF-β superfamily such as BMPs. However, their clinical use is limited due to their pleiotropic extraskeletal effects. Anti-Müllerian Hormone (AMH) is a member of the TGF-β superfamily whose role in bone tissues has never been investigated. To date, AMH functions and the expression of its specific receptor are mainly limited to the reproductive organs. Study design, size, duration We performed in vitro studies on Human Osteoblasts (HOb) to evaluate the expression and the functionality of AMH receptor type-2 and investigate the effects of recombinant AMH (rhAMH) exposure on osteogenic gene expression and osteoblast functions. Participants/materials, setting, methods RT-PCR, real-time PCR and Western blotting analyses were performed in Hob cells cultured both in the presence and in the absence of 100 ng/mL rhAMH to evaluate the effect of AMH exposure on osteoblast differentiation. Alizarin red staining was performed after 14 days of treatment with 100 ng/mL rhAMH to assess the mineralized nodule formation. Main results and the role of chance Our study revealed the presence of a strong mRNA and protein expression of AMHRII in HOb cells, thus indicating that osteoblasts may represent a specific target for exogenous AMH treatment. We also observed that rhAMH exposure increased SMAD 1/5 protein levels, demonstrating that the binding of AMHRII by AMH was able to activate the SMAD-mediated AMH signalling pathway. Interestingly, we also observed that AMH treatment upregulates mRNA levels of crucial osteoblastic transcription factors such as RUNX2 and OSX and extracellular matrix proteins including OPN and OC. Consistently, the prolonged exposure to AMH also promoted matrix mineralization in HOb cells, as demonstrated by both increased ALP gene expression and the greater amount of mineralized nodules observed in AMH-treated osteoblasts compared to untreated controls. Limitations, reasons for caution The study was conducted only in HOb cells. Additional functional studies are needed to investigate the action mechanism of AMH in the other bone cells to understand the mechanisms by which AMH may influence bone homeostasis and pave the way for subsequent in vivo studies. Wider implications of the findings This study may have translation value in opening the perspective that AMH may be an effective candidate to counteract bone loss in osteoporotic patients by developing a new therapy selectively targeting osteoblast with minimal off-target effect. Trial registration number Not Applicable

GRT-X Stimulates Dorsal Root Ganglia Axonal Growth in Culture via TSPO and Kv7.2/3 Potassium Channel Activation.

GRT-X, which targets both the mitochondrial translocator protein (TSPO) and the Kv7.2/3 (KCNQ2/3) potassium channels, has been shown to efficiently promote recovery from cervical spine injury. In the present work, we investigate the role of GRT-X and its two targets in the axonal growth of dorsal root ganglion (DRG) neurons. Neurite outgrowth was quantified in DRG explant cultures prepared from wild-type C57BL6/J and TSPO-KO mice. TSPO was pharmacologically targeted with the agonist XBD173 and the Kv7 channels with the activator ICA-27243 and the inhibitor XE991. GRT-X efficiently stimulated DRG axonal growth at 4 and 8 days after its single administration. XBD173 also promoted axonal elongation, but only after 8 days and its repeated administration. In contrast, both ICA27243 and XE991 tended to decrease axonal elongation. In dissociated DRG neuron/Schwann cell co-cultures, GRT-X upregulated the expression of genes associated with axonal growth and myelination. In the TSPO-KO DRG cultures, the stimulatory effect of GRT-X on axonal growth was completely lost. However, GRT-X and XBD173 activated neuronal and Schwann cell gene expression after TSPO knockout, indicating the presence of additional targets warranting further investigation. These findings uncover a key role of the dual mode of action of GRT-X in the axonal elongation of DRG neurons.