A preliminary study on benthic nutrient exchange across sediment-water interfaces in a shallow marine protected area of the Northwestern Arabian Gulf.

Sedimentary processes are expected to play a crucial role in macronutrient cycling of the shallow Arabian Gulf. To investigate this aspect, sediment cores were collected from the shallow intertidal and subtidal expanses of the first Marine Protected Area (MPA) of Kuwait in the Northwestern Arabian Gulf (NAG). Porewater nutrient profiling and whole core incubation experiments were conducted to measure the nutrient fluxes, both with and without the addition of the nitrification inhibitor allylthiourea (ATU). The porewater data confirmed the potential of sediments to host multiple aerobic and anaerobic pathways of nutrient regeneration. The average (±SD) of net nutrient fluxes from several incubation experiments indicated that ammonium (NH4+) predominantly fluxed out of the sediment (3.81 ± 2.53 mmol m-2 d-1), followed by SiO44- (3.07 ± 1.21 mmol m-2 d-1). In contrast, the average PO43- flux was minimal, at only 0.06 ± 0.05 mmol m-2 d-1. Fluxes of NO3- (ranged from 0.07 ± 0.005 to 1.16 ± 0.35 mmol m-2 d-1) and NO2- (0.03 ± 0.003 to 0.71 ± 0.21 mmol m-2 d-1) were moderate, which either reduced or reversed in the presence of ATU (-0.001 ± 0.0001 to 0.01 ± 0.0001 mmol m-2 d-1 and -0.001 ± 0.0003 to 0.006 ± 0.001 mmol m-2 d-1 for NO3- and NO2- respectively). Thus, this study provides preliminary experimental evidence that nitrification can act as a source of NO3- and NO2- as well as contribute towards the relatively high concentrations of NO2- (>1 in the gulf waters.

Aeolian dust and hydro-biological characteristics: Decoding dust storm impacts on phytoplankton in the northern Arabian Gulf.

Aeolian dust is an essential source of growth-limiting nutrients for marine phytoplankton. Despite being at the core of the Global Dust Belt, the response of the Arabian Gulf ecosystem to such atmospheric forcing is rarely documented. Here, the hydro-biological effect of mineral dust was studied in the northern Arabian Gulf (NAG) off Kuwait through monthly water sampling (December 2020 to December 2021), dust-storm follow-up sampling, and mineral dust and nutrient addition in-situ experiments. The multivariate analysis of oceanographic data revealed pronounced hydro-biological seasonality. The mineral dust deposition during two severe dust storm events in March and June 2021 showed a spatially varying effect of dust on coastal waters. The dust storms elevated the surface dissolved iron levels by several magnitudes, increased the dissolved inorganic nitrogen and phosphorous levels, changed their stoichiometry, and offset the hydrobiological seasonality. In the microcosms, dust input temporarily reduced phytoplankton phosphorous limitation in a dose-dependent manner when mesozooplankton (copepods) grazing was minimal. The microphytoplankton response to mineral dust inputs was comparable to that with nitrogen and phosphorous treatment. While Both treatments increased diatom size structure and biomass, the abundance of single-celled diatoms was comparatively higher in dust treatment. Multivariate analysis indicated that dust deposition alters the hydrographical properties of the surface ocean during dust storm events. The effects, though transient, were traceable for 3-16 days post-storm in coastal waters. The response of the summer phytoplankton to these changes, if delayed or muted, should be interpreted with caution given the summer water column stratification, the high nitrogen: phosphorous ratio and the low phosphorous solubility of aerosol dust, and the complex pelagic microbial food web interactions in the NAG. This study thus underlines the importance of a multivariate approach in documenting the ecological implications of Aeolian dust storms on marine environments closer to the dust source regions.

Dynamic hydrographic and water-quality variations in the northwestern Arabian Gulf, a sinking zone of reverse estuarine circulation.

Continuous measurements of hydrographic, hydrodynamic, and water quality showed marked diurnal, tidal, and seasonal variabilities in Kuwait Bay, a stressed coastal system in the northwestern Arabian/Persian Gulf. Advection of water masses and seasonality in vertical mixing regulated the Bay's hydrographic and water quality properties. Intensive stratification in summer had substantial implications on the Bay environment. Kuwait Bay constantly exports dense bottom water laden with dissolved inorganic nutrients and organic matter to the central basin of the Gulf. The export was largest in August under strong water column stratification. These in-situ findings agreed well with earlier studies that corroborated Kuwait Bay as an important area where the phenomenon of reverse estuarine circulation originates in the Gulf. Thus, Kuwait Bay is a significant source of nutrients and organic matter to the Gulf Deep Water that flows into the core of the oxygen minimum zone in the northwestern Indian Ocean.

Environmental impact of a series of flash flood events on a hypersaline subtropical system in the Northwestern Arabian Gulf.

A series of flash floods that swamped urban drainage systems in Kuwait in November-December 2018 drastically altered coastal hydrography. The phytoplankton responded quickly to the nutrient supply from land and reduced salinity despite elevated turbidity, as evident from rapid increases in chlorophyll a and net community production. Microphytoplankton was dominated by diatoms and the photosynthetic ciliate Myrionecta rubra. Both field observations and microcosm experiments suggested that although increased nutrient availability stimulates diatom growth, microzooplankton grazing controls their outbursts. This study revealed that in a hypersaline system similar to the northern Arabian Gulf, extreme events like flash floods have immediate but short-lived corollaries on coastal phytoplankton due to synergistic effects of bottom-up and top-down factors. The patterns are comparable to those reported from other tropical and sub-tropical systems.

Evaluating the potential area and environmental key factors for Sargassum bed restoration in highly turbid waters of the northwestern Arabian Gulf.

This study evaluates the potential area and the key environmental factors supporting Sargassum bed restoration (SBR) in the highly turbid northwestern Arabian Gulf where rapid coastal development impinges on the marine ecosystem functioning. Water depth was a primary environmental factor governing the distribution of the subtidal macroalgae beds in these turbid waters. The relationship between Sargassum coverage and water depth measured by an echo sounder indicated optimal water depths where the maximum coverage was observed. The availability of stable hard substrate was another key factor introducing heterogeneity in Sargassum coverage. Potential area for the SBR estimated based on the optimal depths was 4.26 km2, whereas only 50% of the potential area (2.19 km2) was currently vegetated due to the absence of hard substrate. The outcomes of this study offer beneficial information toward implementation of the SBR as a part of mitigation measures in future coastal development plans in the region.

Heterotrophic consumption may mask increasing primary production fuelled by anthropogenic nutrient loading in the northern Arabian/Persian Gulf.

Monthly measurements of nitrate, nitrite, ammonium and phosphate at three stations off Kuwait during 2002-2015 revealed considerable inter-annual variability, broadly corresponding to fluctuations in the Shatt-al-Arab River discharge, but a lack of secular increasing trend. Nutrient enrichment experiments during two seasons revealed nitrate uptake, chlorophyll build-up and growth of micro-phytoplankton, even in the presence of ammonium, provided the availability of phosphate. Primary production was mostly nitrogen limited, but anthropogenic nitrogen supply may eventually make it phosphorus limited, especially in summer and in the open Gulf. Anthropogenic nutrient inputs appear to have enhanced biological productivity of the northern Gulf, but heterotrophic consumption, indicated by high respiration rates, probably prevented accumulation of phytoplankton biomass, accounting for the observed lack of chlorophyll increase over the past three decades. Consequently high total organic carbon and emerging hypoxia in the Gulf may lead to expansion/intensification of the oxygen minimum zone of the Arabian Sea.

Does microhabitat influence the testate amoebae communities in the Nameri National Park, northeastern India? A tropical forest perspective.

The community structure of testate amoebae inhabiting different microhabitats (soil and tree-moss) within a tropical forest biome in Nameri National Park, northeastern India, was investigated. A total of 33 testate amoebae species belonging to 13 genera were identified. Species belonging to the class Lobosea constituted 73% of total testate amoebae density in the soil habitat, whereas the class Filosea constituted the most dominant forms (58%) in the moist tree-moss habitat. The relative abundance of species was higher in the tree-moss habitat compared to the soil habitats of the forest. Although multivariate analysis suggested a significant difference in assemblage patterns between the habitats, the turnover in species (i.e., beta diversity) was insignificant. Species accumulation curves (SAC) constructed using both parametric and non-parametric species richness estimators revealed that the asymptote of species richness was achieved by a low number of sample replicates in both habitats. The temperature and pH of the substratum on testate amoebae distribution patterns suggest the importance of additional background factors on testate amoebae community structure. Further studies involving more biotopes, seasons, and trophic interactions are recommended to document a complete record of testate amoebae diversity and their interactions with environmental gradients in the tropical forest biomes of northeastern India.

Environmental characterization of a semiarid hyper saline system based on dissolved trace metal-macronutrient synergy: A multivariate spatio-temporal approach.

Trace metals and macronutrients play key roles in marine biogeochemical processes. Despite their higher availability, no reliable information is available on their ecological role in the hypersaline waters of NW Arabian Gulf. The present study identifies their synergistic effect on environmental characteristics and autotrophic biomass in the shallow coastal and offshore waters, off Kuwait on a seasonal basis. Surface water samples collected along four predefined transects were analyzed for physico-chemical and biological variables during summer (n=27) and winter (n=27) seasons. Multivariate analyses revealed clear spatial and seasonal trends, and identified the best suit of environmental variables responsible for the seasonal variability in phytoplankton biomass. Using statistically derived supply to demand ratios, carbon-trace metal stoichiometry, and bioavailability of trace metals, we propose possible growth regulating factors for phytoplankton on a seasonal basis in the NW Arabian Gulf, off Kuwait. This is the first report on trace metals from the entire Arabian Gulf, using clean techniques and multivariate statistical approach.

A TLR4-interacting SPA4 peptide inhibits LPS-induced lung inflammation.

The interaction between surfactant protein-A (SP-A) and TLR4 is important for host defense. We have recently identified an SPA4 peptide region from the interface of SP-A-TLR4 complex. Here, we studied the involvement of the SPA4 peptide region in SP-A-TLR4 interaction using a two-hybrid system, and biological effects of SPA4 peptide in cell systems and a mouse model. HEK293 cells were transfected with plasmid DNAs encoding SP-A or a SP-A-mutant lacking SPA4 peptide region and TLR4. Luciferase activity was measured as the end-point of SP-A-TLR4 interaction. NF-κB activity was also assessed simultaneously. Next, the dendritic cells or mice were challenged with Escherichia coli-derived LPS and treated with SPA4 peptide. Endotoxic shock-like symptoms and inflammatory parameters (TNF-α, NF-κB, leukocyte influx) were assessed. Our results reveal that the SPA4 peptide region contributes to the SP-A-TLR4 interaction and inhibits the LPS-induced NF-κB activity and TNF-α. We also observed that the SPA4 peptide inhibits LPS-induced expression of TNF-α, nuclear localization of NF-κB-p65 and cell influx, and alleviates the endotoxic shock-like symptoms in a mouse model. Our results suggest that the anti-inflammatory activity of the SPA4 peptide through its binding to TLR4 can be of therapeutic benefit.

Surfactant protein-A and toll-like receptor-4 modulate immune functions of preterm baboon lung dendritic cell precursor cells.

Lung infections are important risk factors for an increased morbidity and mortality in prematurely-delivered babies. Immaturity of the innate immune components makes them extremely susceptible to infection. Recently, we isolated lung dendritic cell (DC)-precursor cells from preterm fetal baboons. The isolated cells were found to be defective in phagocytosing Escherichia coli under basal conditions. In this study, we investigated the effects of exogenously-added purified native lung surfactant protein (SP)-A and recombinant toll-like receptor (TLR)-4-MD2 proteins on phagocytic uptake and cytokine secreting ability of fetal baboon lung DC-precursor cells. The cells were pulsed with SP-A and/or TLR4-MD2 proteins and the phagocytic function was investigated by incubating the cells with fluorescent-labeled E. coli bioparticles and analyzed by spectrofluorometry. The amounts of TNF-α secreted in cell-free supernatants were measured by ELISA. Our results demonstrate that SP-A and TLR4-MD2 proteins, whether added alone or together, induce phagocytosis of E. coli (p<0.05). The SP-A does not affect TNF-α secretion, while the TLR4-MD2 protein induces TNF-α. However, simultaneous addition of SP-A with TLR4-MD2 protein reduces the TLR4-MD2-protein induced TNF-α to basal level. In conclusion, our results indicate that an exogenous administration of SP-A can potentially induce phagocytic activity and anti-inflammatory effect in preterm babies, and help control infection and inflammation.

Curcumin regulates low-linear energy transfer γ-radiation-induced NFκB-dependent telomerase activity in human neuroblastoma cells.

PURPOSE: We recently reported that curcumin attenuates ionizing radiation (IR)-induced survival signaling and proliferation in human neuroblastoma cells. Also, in the endothelial system, we have demonstrated that NFκB regulates IR-induced telomerase activity (TA). Accordingly, we investigated the effect of curcumin in inhibiting IR-induced NFκB-dependent hTERT transcription, TA, and cell survival in neuroblastoma cells. METHODS AND MATERIALS: SK-N-MC or SH-SY5Y cells exposed to IR and treated with curcumin (10-100 nM) with or without IR were harvested after 1 h through 24 h. NFκB-dependent regulation was investigated either by luciferase reporter assays using pNFκB-, pGL3-354-, pGL3-347-, or pUSE-IκBα-Luc, p50/p65, or RelA siRNA-transfected cells. NFκB activity was analyzed using an electrophoretic mobility shift assay and hTERT expression using the quantitative polymerase chain reaction. TA was determined using the telomerase repeat amplification protocol assay and cell survival using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltertrazolium bromide and clonogenic assay. RESULTS: Curcumin profoundly inhibited IR-induced NFκB. Consequently, curcumin significantly inhibited IR-induced TA and hTERT mRNA at all points investigated. Furthermore, IR-induced TA is regulated at the transcriptional level by triggering telomerase reverse transcriptase (TERT) promoter activation. Moreover, NFκB becomes functionally activated after IR and mediates TA upregulation by binding to the κB-binding region in the promoter region of the TERT gene. Consistently, elimination of the NFκB-recognition site on the telomerase promoter or inhibition of NFκB by the IκBα mutant compromises IR-induced telomerase promoter activation. Significantly, curcumin inhibited IR-induced TERT transcription. Consequently, curcumin inhibited hTERT mRNA and TA in NFκB overexpressed cells. Furthermore, curcumin enhanced the IR-induced inhibition of cell survival. CONCLUSIONS: These results strongly suggest that curcumin inhibits IR-induced TA in an NFκB dependent manner in human neuroblastoma cells.

Effect of black raspberry extract in inhibiting NFkappa B dependent radioprotection in human breast cancer cells.

Black raspberry extracts (RSE) have been shown to inhibit cancer cell growth and stimulate apoptosis. Also, studies have demonstrated that RSE inhibits transcriptional regulators including NFkappa B. Accordingly, we investigated the effect of RSE in inhibiting radiation (IR) induced NFkappa B mediated radioprotection in breast adenocarcinoma cells. MCF-7 cells were exposed to IR (2Gy), treated with RSE (0.5, 1.0, 2.0 micro g/ml) or treated with RSE (1.0 micro g/ml) followed by IR exposure, and harvested after 1, 3, 6, 24, 48, and 72 h. NFkappa B DNA-binding activity was measured by EMSA and phosphorylated Ikappa Balpha by immunoblotting. Expression of IAP1, IAP2, XIAP and survivin were measured by QPCR and immunoblotting. Cell survival was measured using MTT assay and cell death using Caspase-3/7 activity. Effect of RSE on IR induced MnSOD, TNFalpha, IL-1alpha and MnSOD activity was also determined. RSE inhibited NFkappa B activity in a dose-dependent manner. Also, RSE inhibited IR-induced sustained activation of NFkappa B, and NFkappa B regulated IAP1, IAP2, XIAP, and survivin. In addition, RSE inhibited IR-induced TNFalpha, IL-1alpha, and MnSOD levels and MnSOD activity. RSE suppressed cell survival and enhanced cell death. These results suggest that RSE may act as a potent radiosensitizer by overcoming the effects of NFkappa B mediated radioprotection in human breast cancer cells.

NFkappaB signaling related molecular alterations in human neuroblastoma cells after fractionated irradiation.

Radiotherapy has been used as an adjunctive local-control modality for high-risk neuroblastoma. However, relapse due to radioresistance affects the success of radiotherapy. Ascertaining the fractionated radiation (FIR) modulated molecular targets is imperative in targeted molecular therapy. Accordingly, we investigated the (i) expression of genes representing six functional pathways; (ii) NFkappaB DNA-binding activity and (iii) expression of radioresponsive molecules after single dose (10 Gy) radiation (SDR) and FIR (2 Gy x 5) in human neuroblastoma cells. Alterations in gene expression were analyzed using QPCR-profiling, NFkappaB activity using electrophoretic mobility shift assay (EMSA) and pIkappaBalpha using immunoblotting. Modulations in TNFalpha, IL-1alpha, pAKT, IAP1, IAP2, XIAP, survivin, MnSOD, BID, Bak, MyD88 and Vegfc were determined using quantitative real-time PCR (Q-PCR) and immunoblotting. Compared to SDR, FIR significantly induced the expression of 25 genes and completely suppressed another 30 genes. Furthermore, FIR induced NFkappaB-DNA-binding activity and IkappaBalpha phosphorylation. Similarly, we observed an induced expression of IAP1, IAP2, XIAP, Survivin, IL-1alpha, MnSOD, Bid, Bak, MyD88, TNFalpha and pAKT in cells exposed to FIR. The results of the study clearly show distinct differences in the molecular response of cells between SDR and FIR. We identified several potential targets confining to NFkappaB signaling cascade that may affect radio-resistance after FIR.

NFkappaB activity and transcriptional responses in human breast adenocarcinoma cells after single and fractionated irradiation.

Radiotherapy is considered mandatory for breast cancer patients undergoing conservative surgery and for women at high risk of recurrence. However, relapse due to radio-resistance affects the success of radiotherapy. Ascertaining the fractionated radiation (FIR) modulated molecular targets is important to make tumors more susceptible to molecular targeted therapy. Accordingly, we investigated the (i) expression of 84 genes representing six functional pathways; (ii) NFkappaB DNA binding activity and; (iii) expression of radio-responsive molecules after single dose (10Gy) radiation (SDR) and FIR (2Gyx5). MCF-7 cells exposed to SDR or FIR were analyzed for alterations in gene expression using QPCR-profiling. NFkappaB DNA binding activity was analyzed using EMSA and pIkappaB using immunoblotting. Expression of TNFalpha, IL-1alpha, pAKT, IAP1, IAP2, XIAP, survivin, MnSOD, BID and Bak were determined using QPCR and/or immunoblotting. Compared to SDR, FIR significantly induced 60 genes and completely suppressed 14 genes. Furthermore, FIR induced NFkappaB-DNA binding activity and IkappaBalpha phosphorylation. Like-wise, FIR induced the expression of IAP1, IAP2, XIAP Survivin, MnSOD, TNFalpha, pAKT and IL-1alpha. The results of the study clearly show distinct differences in the molecular response of cells between SDR and FIR exposures. We identified several potential targets that may affect radio-resistance following FIR.

Curcumin inhibits NFkappaB mediated radioprotection and modulate apoptosis related genes in human neuroblastoma cells.

Curcumin has been shown to exhibit growth inhibitory effects and induce apoptosis in a broad range of tumors. Accordingly, we investigated the radiosensitizing effects of curcumin in human neuroblastoma cells. SK-N-MC cells exposed to either 2 Gy alone, or pretreated with curcumin (100 nM) or NFkappaB inhibitor peptide SN50 (50 nM) and exposed to 2 Gy were harvested after 48 h. Radioresistance was measured using clonogenic and MTT assay, NFkappaB DNA-binding activity using electrophoretic mobility shift assay, and apoptosis using Annexin V-FITC staining. Pathway (apoptosis) specific microarrays were used to measure gene expression and validated using QPCR. Radiation markedly enhanced the NFkappaB DNA-binding activity. Pre-treating the cells either with curcumin or SN50 significantly suppressed the radiation induced NFkappaB. Also, curcumin or SN50 pretreatment enhanced the radiation induced inhibition of cell survival. Microarray analysis revealed that curcumin enhanced the radiation induced activation of caspases, other pro-apoptotic and death effector molecules and, inhibit anti-apoptotic/survival molecules. In addition, curcumin markedly suppressed the radiation induced TNF super family genes. These results suggest that curcumin is a potent radiosensitizer and may act by overcoming the effects of radiation-induced NFkappaB mediated pro-survival gene expression in neuroblastoma.

Alteration of apoptotic signaling molecules as a function of time after radiation in human neuroblastoma cells.

Ascertaining the time-dependent regulation of induced apoptosis and radioresistance is important to understand the relationship between the level of spontaneous apoptosis in cells and their radiosensitivity. Accordingly, we investigated the time-dependent expression of apoptosis related genes and radioresistance in neuroblastoma cells. Serum-starved human SK-N-MC cells were exposed to low linear energy transfer (LET) radiation (2 Gy) and incubated for 15, 30, 45 min, and 48 h. Radioresistance was investigated by examining the NF kappa B DNA-binding activity, cellular toxicity, DNA fragmentation, and expression of apoptotic signal transduction molecules. NF kappa B DNA binding activity was analyzed using electrophoretic mobility shift assay (EMSA). Cellular toxicity was measured using MTT assay. DNA fragmentation was quantified by labeling with fluorescein-conjugated deoxynucleotides. Microarray analysis was performed using cDNA microarray and relative gene expression was measured as % GAPDH and, subsequently validated using Q-PCR. Induction of NF kappa B analyzed using EMSA showed an increased DNA-binding activity at all time points investigated. Induced DNA fragmentation was observed after 15, 30, and 45 min post-radiation. Relatively, induced fragmentation was reduced after 48 h. Compared to the untreated controls cellular toxicity was induced with low LET radiation after 15, 30, and 45 min. Conversely, cytotoxicity was relatively less at 48 h after low LET radiation. Microarray analysis after low LET radiation revealed time-dependent modulation of apoptosis-related genes that are involved in radio-adaptation, spontaneous apoptosis-related early-responsive genes and late response genes. These results suggest that the time-dependent regulation of apoptotic response may determine the relationship between the level of spontaneous apoptosis in cells and their radiosensitivity.