Quantifying Biomolecular Interactions Using Slow Mixing Mode (SLOMO) Nanoflow ESI-MS.

Electrospray ionization mass spectrometry (ESI-MS) is a powerful label-free assay for detecting noncovalent biomolecular complexes and is increasingly used to quantify binding thermochemistry. A common assumption made in ESI-MS affinity measurements is that the relative ion signals of free and bound species quantitatively reflect their relative concentrations in solution. However, this is valid only when the interacting species and their complexes have similar ESI-MS response factors (RFs).

Perturbing dimer interactions and allosteric communication modulates the immunosuppressive activity of human galectin-7.

The design of allosteric modulators to control protein function is a key objective in drug discovery programs. Altering functionally essential allosteric residue networks provides unique protein family subtype specificity, minimizes unwanted off-target effects, and helps avert resistance acquisition typically plaguing drugs that target orthosteric sites. In this work, we used protein engineering and dimer interface mutations to positively and negatively modulate the immunosuppressive activity of the proapoptotic human galectin-7 (GAL-7).

T lymphocyte-proliferative responses of harbor seal (Phoca vitulina) peripheral blood mononuclear cells (PBMCs) exposed to pharmaceuticals in vitro.

The ubiquity of pharmaceuticals in the aquatic environment and the accumulation in organisms of lower trophic levels have been documented. The immunotoxicity of these xenobiotics has however been little investigated. This study assessed the effects of pharmaceuticals on the immune responses of harbor seal lymphocytes.

Exposure to low environmental concentrations of manganese, lead, and cadmium alters the serotonin system of blue mussels.

Serotonin plays a crucial role in mussel survival and reproduction. Although the serotonin system can be affected by metals, the effects of environmental concentrations of metals such as manganese (Mn), lead (Pd), and cadmium (Cd) have never been studied in blue mussels. The present study aimed to determine the effects of exposure to Mn, Pb, or Cd on serotonin levels, monoamine oxidase (MAO) activity, and serotonin transporter (SERT) levels in the blue mussel Mytilus edulis.

Dechlorane Plus induces oxidative stress and decreases cyclooxygenase activity in the blue mussel.

Dechlorane Plus (DP) is a chlorinated flame retardant used mainly in electrical wire and cable coating, computer connectors, and plastic roofing materials. Concentrations of DP (syn and anti isomers) are increasingly being reported in aquatic ecosystems worldwide. However, there is exceedingly little information on the exposure-related toxicity of DP in aquatic organisms, especially in bivalves.

Human Primary Trophoblast Cell Culture Model to Study the Protective Effects of Melatonin Against Hypoxia/reoxygenation-induced Disruption.

This protocol describes how villous cytotrophoblast cells are isolated from placentas at term by successive enzymatic digestions, followed by density centrifugation, media gradient isolation and immunomagnetic purification. As observed in vivo, mononucleated villous cytotrophoblast cells in primary culture differentiate into multinucleated syncytiotrophoblast cells after 72 hr. Compared to normoxia (8% O2), villous cytotrophoblast cells that undergo hypoxia/reoxygenation (0.

Sex determination in blue mussels: Which method to choose?

Sexing methods of blue mussels are mostly based on the presence or absence of gametes, and do not take into account reproductive cycle stages. Exposure effects can be affected by the sex of mussels, thus the aim of this study is to determine an efficient sex determination protocol taking into account the reproductive cycle stage. Eight mussel sexing methods were compared.

Metabolic and immune impairments induced by the endocrine disruptors benzo[a]pyrene and triclosan in Xenopus tropicalis.

Despite numerous studies suggesting that amphibians are highly sensitive to cumulative anthropogenic stresses, the role played by endocrine disruptors (EDs) in the decline of amphibian populations remains unclear. EDs have been extensively studied in adult amphibians for their capacity to disturb reproduction by interfering with the sexual hormone axis. Here, we studied the in vivo responses of Xenopus tropicalis males exposed to environmentally relevant concentrations of each ED, benzo[a]pyrene (BaP) and triclosan (TCS) alone (10 μg L(-1)) or a mixture of the two (10 μg L(-1) each) over a 24 h exposure period by following the modulation of the transcription of key genes involved in metabolic, sexual and immunity processes and the cellular changes in liver, spleen and testis.

Effects of a municipal effluent on the freshwater mussel Elliptio complanata following challenge with Vibrio anguillarum.

The purpose of this study was to examine the cumulative effects of exposure to a pathogenic bacteria and municipal effluent in the freshwater mussel Elliptio complanata. Mussels were exposed to increasing concentrations of an ozone-treated effluent at 15°C for 7days. A sub-group of mussels was inoculated with Vibrio anguillarum and exposed to the same conditions as above.

Genotoxic and immunotoxic potential effects of selected psychotropic drugs and antibiotics on blue mussel (Mytilus edulis) hemocytes.

The potential toxicity of pharmaceuticals towards aquatic invertebrates is still poorly understood and sometimes controversial. This study aims to document the in vitro genotoxicity and immunotoxicity of psychotropic drugs and antibiotics on Mytilus edulis. Mussel hemocytes were exposed to fluoxetine, paroxetine, venlafaxine, carbamazepine, sulfamethoxazole, trimethoprim and erythromycin, at concentrations ranging from μg/L to mg/L.

Responses of freshwater mussel (Elliptio complanata) hemocytes exposed in vitro to crude extracts of Microcystis aeruginosa and Lyngbya wollei.

Lyngbya wollei is a benthic filamentous cyanobacterium that produces a toxin analogous to the neurotoxic saxitoxin known as lyngbyatoxin (LYNGTX). Microcystis aeruginosa form blooms in the pelagic area of eutrophic lakes and produce a series of potent hepatotoxins-microcystins (MCYST). The aim of this study in vitro study was to examine the difference between the crude extracts of either M.

In vitro immunotoxicity of untreated and treated urban wastewaters using various treatment processes to rainbow trout leucocytes.

Municipal effluents are known to impede the immune system of aquatic organisms. The purpose of this study was to examine the immunotoxicity of urban wastewaters before and after 6 treatment processes from 12 cities toward trout leucocytes. Freshly prepared trout leucocytes were exposed to increasing concentrations of solid phase (C18) extracts of wastewaters for 24 hr at 150C.

In vitro immunotoxicology of quantum dots and comparison with dissolved cadmium and tellurium.

The increasing use of products derived from nanotechnology has raised concerns about their potential toxicity, especially at the immunocompetence level in organisms. This study compared the immunotoxicity of cadmium sulfate/cadmium telluride (CdS/Cd-Te) mixture quantum dots (QDs) and their dissolved components, cadmium chloride (CdCl2 )/sodium telluride (NaTeO3 ) salts, and a CdCl2 /NaTeO3 mixture on four animal models commonly used in risk assessment studies: one bivalve (Mytilus edulis), one fish (Oncorhynchus mykiss), and two mammals (mice and humans). Our results of viability and phagocytosis biomarkers revealed that QDs were more toxic than dissolved metals for blue mussels.

Energy status and immune system alterations in Elliptio complanata after ingestion of cyanobacteria Anabaena flos-aquae.

Cyanobacteria have often been described as nutritionally poor for herbivorous organisms. To gain additional information on the potential impacts of invertebrates feeding on cyanobacteria, we fed Elliptio complanata mussels with two types of algae: Anabaena flos-aquae (cyanobacteria) and Pseudokirchneriella subcapitata (green algae). Physiological parameters were examined at the energy status, immune system and oxidative stress levels.

Immunotoxic potential of aeration lagoon effluents for the treatment of domestic and hospital wastewaters in the freshwater mussel Elliptio complanata.

Municipal wastewaters are major sources of pollution for the aquatic biota. The purpose of this study was to determine the levels of some pharmaceutical products and the immunotoxic potential of a municipal wastewater aeration lagoon for the treatment of the domestic wastewaters of a small town with wastewater inputs from a 400-bed hospital complex. Endemic mussels were collected, caged and placed in the final aeration lagoon and at sites 1 km upstream and 1 km downstream of the effluent outfall in the receiving river for a period of 14 days.

Proteomic comparison of detergent-extracted sperm proteins from bulls with different fertility indexes.

Intrinsic factors such as proteins modulate the fertilising ability of male gametes. We compared detergent-extracted sperm protein composition of bulls with different fertility indexes in order to highlight putative fertility markers of sperm. Frozen semen from 23 Holstein bulls with documented fertility was used.

Localization of Hsp60 and Grp78 in the human testis, epididymis and mature spermatozoa.

Molecular chaperones of the heat shock proteins (HSP) family are important in numerous cellular processes. In this study, the expression of Hsp60 and Grp78 proteins was investigated in the male reproductive tract. The cellular distribution of Hsp60 and Grp78 proteins was analysed in the human testis and epididymis by immunohistochemical approaches.

Multiple experimental approaches of immunotoxic effects of mercury chloride in the blue mussel, Mytilus edulis, through in vivo, in tubo and in vitro exposures.

Biological impairments due to mercury discharge into the environment are now an issue of global concern. From the three forms of mercury found in aquatic ecosystems, the immunotoxic effects of mercury chloride were examined in the model animal, the blue mussel. In order to investigate the toxic potency of this chemical, three exposure regimes were carried out: chronic exposure of groups of individuals, a new protocol "in tubo" designed for sub-acute exposures of individuals, and acute exposures of target cells.