Mesophotic Zone Research Articles

Insights into the occurrence of phylosymbiosis and co-phylogeny in the holobionts of octocorals from the Mediterranean Sea and Red Sea

BackgroundCorals are the foundational species of coral reefs and coralligenous ecosystems. Their success has been linked to symbioses with microorganisms, and a coral host and its symbionts are therefore considered a single entity, called the holobiont. This suggests that there may be evolutionary links between corals and their microbiomes. While there is evidence of phylosymbiosis in scleractinian hexacorals, little is known about the holobionts of Alcyonacean octocorals.Results16S rRNA gene amplicon sequencing revealed differences in the diversity and composition of bacterial communities associated with octocorals collected from the mesophotic zones of the Mediterranean and Red Seas. The low diversity and consistent dominance of Endozoicomonadaceae and/or Spirochaetaceae in the bacterial communities of Mediterranean octocorals suggest that these corals may have a shared evolutionary history with their microbiota. Phylosymbiotic signals were indeed detected and cophylogeny in associations between several bacterial strains, particularly those belonging to Endozoicomonadaceae or Spirochaetaceae, and coral species were identified. Conversely, phylosymbiotic patterns were not evident in Red Sea octocorals, likely due to the high bacterial taxonomic diversity in their microbiota, but cophylogeny in associations between certain coral and bacterial species was observed. Noteworthy were the associations with Endozoicomonadaceae, suggesting a plausible evolutionary link that warrants further investigations to uncover potential underlying patterns.ConclusionsOverall, our findings emphasize the importance of Endozoicomonadaceae and Spirochaetaceae in coral symbiosis and the significance of exploring host-microbiome interactions in mesophotic ecosystems for a comprehensive understanding of coral-microbiome evolutionary history.

Beautoide A, an Anti-Osteoclastogenic Sterol from Beauveria sp. NBUF147 Associated with an Irciniidae Sponge from the Marine Mesophotic Zone.

Mesophotic coral ecosystems (MCEs), located at depths ranging from 30-150 m, host some of the most diverse yet least explored marine bioresources, particularly significant for the discovery of new bioactive molecules. The fungus Beauveria sp. NBUF147, associated with an Irciniidae sponge from the mesophotic zone at a depth of 82 m, underwent chemical investigation that led to the identification of one new sterol, beautoide A (1), and one reported sterol, 3β,5α,9α-trihydroxy-(22E,24R)-ergosta-7,22-dien-6-one (2). Their structures were determined from analysis of spectroscopic data and X-ray crystallography. Evaluation of biological activity in prednisolone-induced osteoporotic zebrafish showed that 1 was anti-osteoclastogenic in vivo at 3.0 μM.

First Record of Bramble Sharks, Echinorhinus brucus (Echinorhiniformes, Echinorhinidae), in the United Arab Emirates

The first record of bramble sharks (Echinorhinus brucus) in the United Arab Emirates is presented. In situ observations of multiple bramble sharks were made at depths between 460 and 720 m from two piloted submersibles and a remotely operated vessel, representing the first known observations of this species in its native deep-water habitat in Arabia and the Indian Ocean. Notably, this research expands on the documented regional distribution of E. brucus for the Gulf of Oman/Arabian Sea and extends this species’ regional records to deeper mesophotic zones. These findings underscore the need for further research to understand the ecology and distribution of this cryptic shark species, particularly given its global endangered status and the limited knowledge of its regional population dynamics.

Diversity and paleoecological significance of zooxanthellate corals of Oligocene Qom Formation, SE Iran

Zooxanthellate corals are Iran's most abundant macrofossils in the Oligocene-early Miocene Qom Formation deposits in the NE margin of the Tethyan Seaway. Yet, basic knowledge about the diversity of these organisms and their paleoecology is still scarce. This paper uses late Rupelian–Chattian coral data from three exposed sections in the northwestern Jazmurian Lake, SE Iran, as the southeasternmost outcrops of the Qom Formation, are analyzed in terms of diversity and paleoecology. The coralline red algae and larger benthic foraminiferal assemblages associated with the corals have also been used to infer the paleoenvironmental and paleoecological conditions. The following coral genera were recognized: Leptoria, Caulastraea, Hydnophora, Astreopora, Plesiastrea, Thegioastraea, Porites, Goniopora, Platycoenia, Acropora, Tarbellastraea, Favites, Heliastraea, Stylocoenia, Ceratotrochus. The high abundance of z-corals, larger benthic foraminifera (LBF), and coralline red algae demonstrate that the deposition took place in tropical-subtropical warm waters mainly within the euphotic to the mesophotic zones. The presence of various coral fabrics including pillarstone, domestone, and rudstone allowed to infer high to moderate hydrodynamic energy in the studied sections. As well, given the interactions among corals, filter-feeders, green and red algae, bioeroders, and the surrounding environment, it seems that oligotrophic to slightly mesotrophic conditions also had prevailed in the studied area.

A Bajocian event of global coral reef growth: Record from northern Switzerland

Following the extinction event at the Triassic – Jurassic boundary and during the Pliensbachian – Toarcian events, coral reef builders were severely affected, and the number of reef domains during the early Middle Jurassic was at an all-time low. However, new and updated data seem to show that corals were more widespread than previously thought, particularly during the Bajocian (170–168 Ma), but biostratigraphic markers are lacking to constrain the precise timing of reef evolution of this interval. A recently discovered 40 m-thick carbonate unit in northern Switzerland, on the northern Tethys margin, documents this rapid growth. Two cores from this unit display a reef with a high number of specimen but a low coral diversity, dated as Bajocian, which likely grew at a considerable water depth (upper limit of the mesophotic zone or 20–30 m). Additionally, it developed on top of clay mineral-rich accumulation characterised by low relief. Twelve different coral genera were identified, with a strong predominance of Periseris, Isastrea, Thamnasteria and Dendraraea. In the absence of other biostratigraphic markers, palynomorphs were used to provide accurate dating, which shows that palynology is a powerful tool for dating reef deposits. The results show reef growth during the early Bajocian, in the Sauzei and Humphriesianum ammonite zones, based on the palynological data, yielding similar ages to other Bajocian reef localities in the Eastern Paris Basin in France and Luxembourg dated simultaneously. These results indicate that the Middle Jurassic and more specifically the last two ammonite zones of the early Bajocian were periods suitable for coral reef builders, and that global regrowth of coral reefs occurred more rapidly than expected. A new Bajocian reef distribution is presented, showing no reefs at latitudes below 15°. This distribution pattern is probably due to the fact that water temperature was too elevated (> 25 °C) towards the equator for corals, which prevented reef growth. This would tend to favour a scenario in which the global climate, in particular temperature, is a determining factor for coral reef growth.

Mesophotic zone as buffer for biodiversity protection: A promising opportunity to enhance MPA effectiveness

Coastal areas conservation strategies often left deeper habitats, such as mesophotic ones, unprotected and exposed to anthropogenic activities. In this context, an approach for including the mesophotic zone inside protection plans is proposed, considering 27 Italian Marine Protected Areas (MPAs) as a model. MPAs were classified considering their bathymetries, exposure to marine heat waves (MHWs), mass mortality events (MMEs) and, using a local ecological knowledge (LEK) approach, the estimated resilience of certain sessile species after MMEs. Only 8 MPAs contained considerable mesophotic areas, with stronger MHWs mainly occurring in shallower MPAs, and MMEs mostly affecting coralligenous assemblages. Even with only a 10% response rate, the LEK approach provided useful information on the resilience of certain species, allowing us to suggest that the presence of nearby mesophotic areas can help shallower habitats facing climate change, thus making the “deep refugia” hypothesis, usually related to tropical habitats, applicable also for the Mediterranean Sea.

Diverse and ecologically unique mesophotic coral ecosystems in the central Indian Ocean

Research on Mesophotic Coral Ecosystems (MCEs; 30–150 m) has grown exponentially in the last few decades, highlighting their rich diversity and extensive distribution. However, they are still largely underexplored compared to shallow-water coral reefs and frequently remain under-protected and under-represented in marine spatial planning. One reason for the imbalance between the high ecological value of MCEs and the limited levels of protection may be that baseline data on MCEs are largely missing to date, yet are crucial to provide evidence-based information for management actions. Here, we present data on the alpha and beta diversity of the benthic communities within MCEs in the Chagos Archipelago, Indian Ocean. Using imagery collected from Remotely Operated Vehicle surveys, benthic invertebrate megafauna were surveyed along the entire depth gradient from shallow to lower mesophotic depths (15–160 m). The diversity of the benthic communities decreased with increasing depth, from shallow water to the lower mesophotic zone. Nevertheless, the deepest parts of MCEs in the Archipelago displayed higher species richness values than several other shallow subtropical regions. In addition, the benthic communities showed high dissimilarity along the depth gradient, indicating that the key driver of community composition change with depth is species turnover (species replacement), revealing the uniqueness of MCEs. This study presents novel findings on MCEs in the central Indian Ocean, demonstrating that they host a high and unique benthic diversity, and highlighting the need to protect these ecosystems to preserve the overall biodiversity of coral reefs.

Seasonal dynamics of Mediterranean fish communities revealed by eDNA: Contrasting compositions across depths and Marine Fully Protected Area boundaries

Marine fish communities suffer from anthropogenic pressures and climate change, which influence their spatio-temporal dynamics. Marine Protected Areas (MPAs) have been established worldwide to preserve these communities, while mesophotic ecosystems could provide natural refugia. Assessing the extent to which MPAs and deeper ecosystems can mitigate human and climate change impacts requires regular monitoring of temporal community dynamics. Environmental DNA (eDNA) surveys – being time- and cost-effective – can provide valuable insights on biodiversity change. Here, we initiated a long-term study based on eDNA monitoring in an MPA in the north-western Mediterranean Sea that includes areas with various protection levels. Specifically, from June 2021 to January 2023, we collected eDNA samples during the summer, fall, and winter seasons from shallow water (20 m depth), at 40 m depth, and from the mesophotic zone (80 m depth) in a Fully Protected Area (FPA) and in a nearby Lightly Protected Area (LPA) in the Riou archipelago (France). In this short period and relatively small area, we detected a total of 113 actinopterygian and chondrichthyan taxa. Species with high fishing vulnerability had higher detection rates in the FPA than in the LPA, suggesting a positive impact of FPAs on the conservation of these threatened species. A marked seasonal signal in species detections, including significantly lower detections of several species in winter, indicated a combined effect of species biological changes and migration behavior. The seasonality trend was stronger in the FPA than in the LPA, indicating that such areas may modify sub-yearly patterns in communities and ecosystem processes. Fish composition was associated with water depth, with marked species dissimilarities between shallow waters and the mesophotic zone, implying that multiple depths should be considered in MPA monitoring to fully capture the response of biodiversity to management. Our results point to the importance of temporal information combined with extensive sampling across depths and protection levels to fully understand the ecological dynamics and structure of coastal fish communities.

The mesophotic zone of the marine protected area Espíritu Santo, Gulf of California, can function as a refuge for the ichthyofauna

Research on mesophotic reefs has increased thanks to technological advances such as remotely operated vehicles (ROV). The aim of this study was to evaluate differences in ichthyofaunal assemblages between shallow (<30 m) and mesophotic (>30 m) reef zones in three sites of the Parque Nacional zona marina del Archipiélago de Espíritu Santo (PNZMAES), a Marine protected area from the Gulf of California. To accomplish this objective, we used 14 video transects recorded with an ROV in October 2019 between 13 and 72 m depth. From these recordings, we determined species richness and abundance to analyze community structure and estimate biomass. A permutational multivariate analysis of variance was used to analyze differences between zones, sites, and their interaction. The abundance and estimated biomass of trophic groups were plotted to describe the trophic structure of the depth zones at PNZMAES. We also analyzed functional structure, functional richness (FRic), functional evenness (FEve), functional divergence (FDiv), functional specialization (FSpe), and functional originality (FOri) by depth zone and video transect based on six biological traits and estimated biomass. The taxonomic and functional indices were the response variables of linear models, while the explanatory variables were in situ temperature, light, and dissolved oxygen. At PNZMAES, 61 species were registered: 42 species in the shallow zone (high abundance and estimated biomass of planktivorous) and 40 species in the mesophotic zone (high abundance of planktivorous and high estimated biomass of piscivores). When comparing the ichthyofaunal community structure, differences were observed in richness, abundance, and estimated biomass associated with the depth zone, site, and the interaction of these variables. An overlap of more than 60% in the functional volume between zones was observed. Higher values of FRic, FDiv, and FSpe in the deep zone indicated that ecological functions were equivalent to those in the shallow zone. Finally, the linear models indicated an effect of environmental variables on the ecological indicators. The mesophotic reefs of PNZMAES are potential refuges for certain species (some of them endangered) and should be considered in the proposal of management strategies to protect them from future anthropogenic threats.

A revision of the genus Wrangelia (Wrangeliaceae, Ceramiales) in Bermuda resolves six new species including W. ryancraigii from the mesophotic zone.

Four species of the genus Wrangelia are presently known from the western Atlantic Ocean: W. argus, W. bicuspidata, W. penicillata, and W. gordoniae, with the first three historically being reported from Bermuda. Morphological and molecular barcode (COI-5P) and phylogenetic analyses used in this study (SSU, LSU, rbcL) indicated eight species groupings of Wrangelia in Bermuda, excluding two of the historically recognized species, retaining only W. argus while adding seven new species, of which six are formally described. What had been historically reported as W. penicillata from Bermuda was shown to be distinct from Mediterranean Sea specimens (type locality) and was shown to be a mixture of W. hesperia sp. nov. and W. incrassata sp. nov. Along with these two, three other new species (W. laxa sp. nov., W. ryancraigii sp. nov., and W. secundiramea sp. nov.) have complete rhizoidal cortication tightly covering axial cells of indeterminate axes below the apices, distinguishing them from the two local incompletely corticated congeners W.argus and W. abscondita sp. nov., the latter a morphologically cryptic sister species with W. bicuspidata from the Caribbean Sea. Only one of the new species, W. ryancraigii, has thus far been observed in the mesophotic zone off the Bermuda platform, and it is morphologically cryptic with the euphotic zone's W. laxa.

Mesoscale Eddies Influence Coral Reef Environments in the Northwest Gulf of Mexico

AbstractCoral reefs globally are experiencing more frequent and severe warming events due to anthropogenic driven climate change. Subtropical reefs experience more seasonal variability than lower latitude reefs making them typically more resilient to climate change. With relatively stable coral cover in comparison to other global coral reefs, Flower Garden Banks National Marine Sanctuary (FGBNMS) in the Gulf of Mexico is a series of 17 reefs and banks located on the continental shelf‐edge containing a variety of shallow (0–30 m) and mesophotic (30–150 m) coral reef habitats. Here, we use satellite data products to associate open ocean Argo float profiles with eddy features over FGBNMS to study the shelf‐edge reef environment spanning nearly two decades (2003–2022). Satellite data show that FGBNMS is frequently influenced (∼15 days/month) by mesoscale eddies. The upper water column variability (0–25 m) is most influenced by the seasonal mixed layer despite eddy interaction. Subsurface seasonal ranges of temperature and salinity are enhanced or suppressed depending on the influence of eddies in relation to the mixed layer depth. Within the mesophotic zone (0–150 m), the largest range of thermal variability between anticyclonic and cyclonic eddies is between 50 and 150 m upwards of 5°C. However, these observed dynamics will likely change as a result of eddy variability associated with projected warming and Loop Current weakening, leading to increased thermal stress in the future.

Hidden from plain sight: Sclerosponges as environmental archives of the ocean conditions from the surface to the mesophotic zone

Hidden from plain sight: Sclerosponges as environmental archives of the ocean conditions from the surface to the mesophotic zone

Expansion of an established fishery-independent survey into the US Virgin Islands' upper mesophotic zone: feasibility and management implications

A three-year pilot study, the Deep Coral Reef Monitoring Program (DCRMP), expanded the National Coral Reef Monitoring Program's (NCRMP) established fishery-independent, diver-based reef fish visual survey to upper mesophotic reefs (&gt;30 to 50 m) in the United States Caribbean for the first time. The new DCRMP sample domain (&gt;30 to 50 m) encompassed 2.4 times more survey area than NCRMP (0 to ≤30 m) and collected high quality data (CV &lt;20%) on coral reef fishes [three survey years, 29 (5) species; mean (standard deviation)]. For the four representative, fishery-targeted, analysis species selected (i. e., grouper, snapper, triggerfish, and parrotfish), domain-wide density and length comparisons between surveys showed similar or statistically higher abundances and larger lengths for fishes at deeper depths (&gt;30 to 50 m). These results highlight the importance of surveying the entire insular shelf in St. Thomas and St. John, US Virgin Islands for fisheries management applications. Furthermore, the DCRMP survey leveraged NCRMP's methods and resources resulting in a seamless extension to deeper waters. However, if these programs were fully integrated and optimized within a single survey design, approximately half the sites would be needed to achieve the same level of precision, offering substantial time and cost savings. The principles of probabilistic sampling successfully used in the present fishery-independent survey design (0 to 50 m) can be applied more broadly to develop an "ideal" large-scale, multi-gear survey from 0 to about 500 m to encompass the entire depth ranges of managed species in the US Caribbean.

Upper mesophotic reef fish assemblages at Bahía de Banderas, Mexico.

There is no information on the species associated with the mesophotic reefs of Banderas Bay, located in the central Mexican Pacific. This study analysed the reef fish assemblage from three depths (50, 60 and 70 m) in three sampling sites of the southern submarine canyon of the Bay: Los Arcos, Bajo de Emirio and Majahuitas. Several analyses were performed to test the hypothesis that there are important differences in fish abundance and species composition between sites and depths. Twenty-two species of bony fishes grouped in 14 families were recorded. PERMANOVA results showed that there were no significant differences in fish diversity parameters between sites, indicating a certain uniformity in their distribution. However, nine species were exclusive to one site and depth (five singleton species with only one individual recorded and four unique species recorded only once). On the other hand, there were significant differences between depths, mainly between 50 and 70 m. Diversity decreases with depth and species composition changes. SIMPER, Shade Plot and NMDS analysis show the most representative species at each depth, with at least half of the species (11) recorded only at 50 m and four species at the deeper levels (60 - 70 m). The observed assemblage includes several of the most caught species in the shallow water artisanal fishery, which is the most traditional and common type of fishery in the Bay. In addition, the Pomacanthuszonipectus (Cortés angelfish) is of particular interest, as it has a special protection status in the official Mexican standard (NOM-059-SEMARNAT, 2010) due to its use as an ornamental species in aquaria. We hypothesised that the mesophotic zone may serve as a refuge for these fishes, so we propose that the information obtained is an important basis for new research aimed at the sustainable management of fisheries in the area.

Mesophotic zone as refuge: acclimation and in-depth proteomic response of yellow gorgonians in the Mediterranean sea

Mesophotic zone as refuge: acclimation and in-depth proteomic response of yellow gorgonians in the Mediterranean sea

Mesophotic corals in Hawai'i maintain autotrophy to survive low-light conditions.

In mesophotic coral ecosystems, reef-building corals and their photosynthetic symbionts can survive with less than 1% of surface irradiance. How depth-specialist corals rely upon autotrophically and heterotrophically derived energy sources across the mesophotic zone remains unclear. We analysed the stable carbon (δ13C) and nitrogen (δ15N) isotope values of a Leptoseris community from the 'Au'au Channel, Maui, Hawai'i (65-125 m) including four coral host species living symbiotically with three algal haplotypes. We characterized the isotope values of hosts and symbionts across species and depth to compare trophic strategies. Symbiont δ13C was consistently 0.5‰ higher than host δ13C at all depths. Mean colony host and symbiont δ15N differed by up to 3.7‰ at shallow depths and converged at deeper depths. These results suggest that both heterotrophy and autotrophy remained integral to colony survival across depth. The increasing similarity between host and symbiont δ15N at deeper depths suggests that nitrogen is more efficiently shared between mesophotic coral hosts and their algal symbionts to sustain autotrophy. Isotopic trends across depth did not generally vary by host species or algal haplotype, suggesting that photosynthesis remains essential to Leptoseris survival and growth despite low light availability in the mesophotic zone.

Irpetones A and B, Anti-Osteoclastic Heptaketides from a Marine Mesophotic Zone Ircinia Sponge-Associated Fungus Irpex sp. NBUF088.

Chemical investigation of Irpex sp. NBUF088, associated with an Ircinia sp. sponge located at an 84 m deep mesophotic zone, led to the discovery of two new heptaketides, named irpetones A (1) and B (2). Their structures were identified by analysis of spectroscopic data and quantum-chemical calculations. Compound 1 exhibited inhibition against the receptor activator of NF-κB ligand-induced osteoclastogenesis in bone marrow monocytes with an IC50 of 6.3 ± 0.2 μM, causing no notable cytotoxicity. It was also determined that 1 inhibited the phosphorylation of ERK1/2-JNK1/2-p38 MAPKs and the nuclear translocation of NF-κB, consequently suppressing the activation of MAPK and NF-κB signaling pathways induced by the NF-κB ligand.

Metabolites with Anti-acetylcholinesterase Activities from Marine Mesophotic Zone Halichondria Sponge-associated Fungus Chaetomium sp. NBUF152

Background: Mesophotic coral ecosystems (MCEs) are light-dependent ecosystems found from 30 to 150 m in depth that are highly diverse and relatively understudied worldwide. Methods: One new indole alkaloid, chaetoid A (1), one known analog, indigodole B (2), and one known β-carboline type alkaloid 1-acetyl-β-carboline (3), along with three known cyclic depsipeptides, turnagainolide A (4), turnagainolide B (5), arthroamide (6)—were isolated from cultures of fungus Chaetomium sp. NBUF152 is associated with a 66 m deep mesophotic zone Halichondria sponge. Their structures were determined by analysis of spectroscopic data (NMR, HRESIMS, UV, IR, optical rotation, and ECD). The in vitro bioassay results showed that compounds 1-6 exhibited moderate inhibition against acetylcholinesterase (AchE). Results: The results showed that these compounds displayed moderate anti-AchE activities with IC50 values of 25.97 μmol/L, 25.80 μmol/L, 21.07 μmol/L, 23.49 μmol/L and 21.28 μmol/L, 25.3 μmol/L. Conclusion: This work could lead to additional investigation into the potential of marine organisms in MCEs as a source for discovering natural products with interesting structures and bioactivities.

Organellar genomic characterization of Anunuuluaehu liula representing a new genus and species of Phyllophoraceae (Gigartinales, Rhodophyta) from the mesophotic zone of Hawai'i.

Over the last 2 decades, routine collections in the Hawaiian Archipelago have expanded to mesophotic reefs, leading to the discovery of a new red algal genus and species, here described as Anunuuluaehu liula gen. et sp. nov. This study provides a detailed genus and species description and characterizes chloroplast and mitochondrial organellar genomes. The new genus, Anunuuluaehu, shares many characteristics with the family Phyllophoraceae and shows close similarities to Archestennogramma and Stenogramma, including habit morphology, nemathecia forming proliferations at the outer cortex with terminal chains of tetrasporangia, and carposporophytes with multi-layered pericarps. The single species in this genus exhibits distinctive features within the Phyllophoraceae: the presence of single-layer construction of large medullary cells and the development of long, tubular gonimoblastic filaments. Multi-gene phylogenetic analyses confirmed it as a unique, monophyletic lineage within the family. Cis-splicing genes, interrupted by intron-encoded proteins within group II introns, are present in both the chloroplast and mitochondrial genomes of A. liula. Notably, a specific region of the coxI group II intron exhibits similarity to fungal introns. Anunuuluaehu liula is presumed to be endemic to the Hawaiian Archipelago and thus far is known to live solely at mesophotic depths from Hōlanikū to Kaho'olawe ranging from 54 to 201 m, which is the deepest collection record of any representative in the family. Overall, this study enhances our understanding of the genomic and taxonomic complexities of red algae in mesophotic habitats, emphasizing the significance of continued research in this area to uncover further insights into evolutionary processes and biogeographic patterns.

Pestanoid A, a Rearranged Pimarane Diterpenoid Osteoclastogenesis Inhibitor from a Marine Mesophotic Zone Chalinidae Sponge-Associated Fungus, Pestalotiopsis sp. NBUF145.

One novel rearranged pimarane diterpenoid, pestanoid A (1), and two reported molecules, nodulisporenones A (2) and B (3), were discovered from Pestalotiopsis sp. NBUF145 fungus associated with a 62 m deep mesophotic ("twilight") zone Chalinidae sponge. The structures of 1-3 were identified by spectrometry, spectroscopy, quantum-chemical calculations, and X-ray crystallography. Compounds 1 and 2 inhibited bone marrow monocyte osteoclastogenesis in vitro with the IC50 values 4.2 ± 0.2 μM and 3.0 ± 0.4 μM, respectively, without observed cytotoxicity. Both 1 and 2 suppressed the receptor activator of NF-kB ligand-induced MAPK and NF-κB signaling by inhibiting the phosphorylation of ERK1/2-JNK1/2-p38 MAPKs and NF-κB nuclear translocation.