SP-8356 inhibits acute lung injury by suppressing inflammatory cytokine production and immune cell infiltration.

This study investigated the anti-inflammatory and protective properties of SP-8356, a synthetic derivative of (1S)-(-)-verbenone, in a mouse model of LPS-induced acute lung injury (ALI). By targeting intracellular signaling pathways and inflammatory responses, SP-8356 demonstrated a potent ability to attenuate deleterious effects of proinflammatory stimuli. Specifically, SP-8356 effectively inhibited the activation of crucial signaling molecules such as NF-κB and Akt, and subsequently dampened the expression of inflammatory cytokines in various lung cellular components. Intervention with SP-8356 treatment also preserved the structural integrity of the epithelial and endothelial barriers. By reducing immune cell infiltration into inflamed lung tissue, SP-8356 exerted a broad protective effect against ALI. These findings position SP-8356 as a promising therapeutic candidate for pulmonary inflammatory diseases that cause ALI.

Validation of reference genes for quantitative real-time PCR in chemical exposed and at different age's brackish water flea Diaphanosoma celebensis.

Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), a primary approach for evaluating gene expression, requires an appropriate normalization strategy to confirm relative gene expression levels by comparison, and rule out variations that might occur in analytical procedures. The best option is to use a reference gene whose expression level is stable across various experimental conditions to compare the mRNA levels of a target gene. However, there is limited information on how the reference gene is differentially expressed at different ages (growth) in small invertebrates with notable changes such as molting. In this study, expression profiles of nine candidate reference genes from the brackish water flea, Diaphanosoma celebensis, were evaluated under diverse exposure to toxicants and according to growth. As a result, four different algorithms showed similar stabilities of genes for chemical exposures in the case of limited conditions using the same developmental stage (H2A was stable, whereas Act was fairly unstable in adults), while the results according to age showed a significantly different pattern in suite of candidate reference genes. This affected the results of genes EcRA and GST, which are involved in development and detoxification mechanisms, respectively. Our finding is the first step towards establishing a standardized real-time qRT-PCR analysis of this environmentally important invertebrate that has potential for aquatic ecotoxicology, particularly in estuarine environments.

Oxidative stress responses in brackish water flea exposed to microcystin-LR and algal bloom waters from Nakdong River, Republic of Korea.

Microcystis blooms and the impact of their toxins, particularly microcystin (MC), in coastal ecosystems is an emerging threat, but the species-specific effects of MC and the potential for bioconcentration are not fully understood. We exposed the brackish water flea, Diaphanosoma celebensis, to MC-LR, which showed antioxidant responses measured at the molecular to enzyme levels but no acute toxicity. We extended our experimental investigation to measure the released MC and its uptake by D. celebensis exposed to river water. In a short-term exposure (48 h) experiment, D. celebensis exposed to water from an algal bloom (approximately 2 µg L-1 MC) assimilated more than 50 pg MC per individual. The significant increase of MCs suggests the potential for the species to accumulate MCs. The dose-dependent increase in the antioxidant response observed in the mRNA levels also showed that D. celebensis exposed to diluted algal bloom waters were affected by toxins from cyanobacteria.

Cloning and molecular characterization of estrogen-related receptor (ERR) and vitellogenin genes in the brackish water flea Diaphanosoma celebensis exposed to bisphenol A and its structural analogues.

Although it has previously been shown that bisphenol (BP) analogues may interfere with the normal hormonal regulation by acting as endocrine disrupting chemicals (EDCs), little information is available on effects of BP analogues in invertebrates, particularly on cladocerans. In the present study, we identified estrogen-related receptors (EER), vitellogenin (VTG), and VTG receptor (VtgR) from the brackish water flea Diaphanosoma celebensis, and examined the effects of BPA and the substitutes, BPF and BPS, in different sublethal concentrations. Gene expression varied with time well matched with brooding, suggesting that DcEER, DcVTG, and DcVtgR play a role in reproduction in D. celebensis. qRT-PCR analysis showed that BPA and its substitutes differently modulated mRNA expressions of DcEER, DcVTG, and DcVtgR, indicating that these compounds adversely affect the normal reproduction-related pathway. This study facilitates better understanding of the molecular mode of action of BP analogues on the reproductive system of D. celebensis.

Acute toxicity of bisphenol A and its structural analogues and transcriptional modulation of the ecdysone-mediated pathway in the brackish water flea Diaphanosoma celebensis.

Bisphenol A (BPA) is a representative endocrine disrupting chemical (EDC) that has estrogenic effects in aquatic animals. In recent years, due to the continuing usage of BPA, its analogues have been developed as alternative substances to replace its use. The molting process is a pivotal point in the development and reproduction of crustaceans. However, studies of the effects of EDCs on molting in crustaceans at the molecular level are scarce. In the present study, we examined the acute toxicity of BPA and its analogues bisphenol F (BPF) and S (BPS) to the brackish water flea Diaphanosoma celebensis. We further identified four ecdysteroid pathway - related genes (cyp314a1, EcRA, EcRB, and USP) in D. celebensis, and investigated the transcriptional modulation of these genes during molting and after exposure to BPA and its analogues for 48 h. Sequencing and phylogenetic analyses revealed that these four genes are highly conserved among arthropods and may be involved in development and reproduction in the adult stage. The mRNA expression patterns of cyp314a1, EcRA and USP were matched with the molting cycle, suggesting that these genes play a role in the molting process in the adult stage in cladocerans. Following relative real-time polymerase chain reaction (RT-PCR) analyses, BPA and its analogues were found to modulate the expression of each of these four genes differently, indicating that these compounds can disrupt the normal endocrine system function of D. celebensis. This study improves our understanding of the molecular mode of action of BPA and its analogues in D. celebensis.