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Cellular responses elicited by cell surface receptors differ according to stimulus strength. We investigated how the high-affinity receptor for immunoglobulin E (IgE) modulates the response of mast cells to a high- or low-affinity stimulus. Both high- and low-affinity stimuli elicited similar receptor phosphorylation; however, differences were observed in receptor cluster size, mobility, distribution, and the cells' effector responses. Low-affinity stimulation increased receptor association with the Src family kinase Fgr and shifted signals from the adapter LAT1 to the related adapter LAT2. LAT1-dependent calcium signals required for mast cell degranulation were dampened, but the role of LAT2 in chemokine production was enhanced, altering immune cell recruitment at the site of inflammation. These findings uncover how receptor discrimination of stimulus strength can be interpreted as distinct in vivo outcomes.

Uncouplers of oxidative phosphorylation dissipate the proton gradient, causing lower ATP production. Bacteria encounter several non-classical uncouplers in the environment, leading to stress-induced adaptations. Here, we addressed the molecular mechanisms responsible for the effects of uncouplers in Escherichia coli. The expression and functions of genes involved in phenotypic antibiotic resistance were studied using three compounds: two strong uncouplers, i.e., Carbonyl cyanide m-chlorophenyl hydrazone (CCCP) and 2,4-Dinitrophenol (DNP), and one moderate uncoupler, i.e., Sodium salicylate (NaSal). Quantitative expression studies demonstrated induction of transcripts encoding marA, soxS and acrB with NaSal and DNP, but not CCCP. Since MarA and SoxS are degraded by the Lon protease, we investigated the roles of Lon using a lon-deficient strain (Δlon). Compared to the wild-type strain, Δlon shows compromised growth upon exposure to NaSal or 2, 4-DNP. This sensitivity is dependent on marA but not rob and soxS. On the other hand, the Δlon strain shows enhanced growth in the presence of CCCP, which is dependent on acrB. Interestingly, NaSal and 2,4-DNP, but not CCCP, induce resistance to antibiotics, such as ciprofloxacin and tetracycline. This study addresses the effects of uncouplers and the roles of genes involved during bacterial growth and phenotypic antibiotic resistance. Strong uncouplers are often used to treat wastewater, and these results shed light on the possible mechanisms by which bacteria respond to uncouplers. Also, the rampant usage of some uncouplers to treat wastewater may lead to the development of antibiotic resistance., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Objective: We evaluated the potential neuro-regenerative effects of the mitochondrial uncoupler 2,4-Dinitrophenol in experimental autoimmune neuritis, an animal model for an acute autoimmune neuropathy., Methods: Experimental autoimmune neuritis was induced in Lewis rats. Different concentrations of 2,4-Dinitrophenol (1 mg/kg, 0.1 mg/kg and 0.01 mg/kg) were applied during the recovery phase of the neuritis (at days 18, 22 and 26) and compared to the vehicle. Any effects were assessed through functional, electrophysiological, and morphological analysis via electron microscopy of all groups at day 30. Additional immune-histochemical analysis of inflammation markers and remyelination of the sciatic nerves were performed for the dosage of 1 mg/kg and control., Results: No enhancement of functional or electrophysiological recovery was observed in all 2,4-Dinitrophenol-treated groups. Cellular inflammation markers of T cells (CD3+) were comparable to control, and an increase of macrophages (IbA1+) invasion in the sciatic nerves was observed. Treatment with 2,4-Dinitrophenol reduced axonal swelling in myelinated and unmyelinated fibers with an increased production of brain-derived neurotrophic factor., Conclusion: Our findings do not support the hypothesis that repurposing of the mitochondrial uncoupler 2,4-Dinitrophenol exerts functionally relevant neuro-regenerative effects in autoimmune neuritis., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Introduction: 2,4-dinitrophenol (DNP) is a mitochondrial toxin sometimes used as a weight loss agent. Reports of fatalities from DNP have been increasing since 2000, suggesting an increase in use. Our understanding of DNP toxicity in humans comes from reports to Poison Control and postmortem analyses, sources that are biased to more extreme presentations. This leads to a gap in our knowledge about the adverse effects of DNP at nonlethal doses. Here we investigate the doses and effects of DNP as reported online., Methods: We analyzed publicly available Internet posts that we collected from 2017-2019. The posts came from anonymous users or users who voluntarily self-identified. We collected data from websites whose terms of use allow for the secondary analysis of data that their users agree to make public. We used natural language processing techniques that we had previously developed to extract doses, effects, and substances mentioned in each post., Results: We collected 1,630 posts across 5 online forums and the Reddit forum r/DNP. The posts were from 1,234 unique usernames. The most commonly reported doses were between 150 to 300 mg each day followed by 300 to 450 mg each day. At those doses, the most reported adverse effects were profuse sweating and fatigue. Reports of thermoregulatory (sweating, feeling hot flashes or flushed), fatigue-related, and neurologically related symptoms were statistically significantly more frequent at reported daily doses greater than 150 mg than doses below 150 mg (post-hoc χ2-test with Bonferroni correction). The effects were judged as plausible by two board-certified medical toxicologists. Triiodothyronine, clenbuterol, testosterone, and trenbolone, an androgenic anabolic steroid were the most significantly co-mentioned substances., Conclusions: Fatigue, increased body temperature, and paresthesias from DNP are reported more frequently at doses greater than 150 mg each day than at doses less than 150 mg each day. Online discussions of DNP frequently mention androgenic anabolic steroids and other weight loss agents., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Abdelati et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Although they are widely used as insecticides, acaricides and fungicides in the agriculture or as raw materials in the dye industry, dinitrophenols (DNPs) are extremely noxious, death cases having been registered. These compounds produce cataracts, lower leucocyte levels, disturb the general metabolism and can cause cancer. It is also assumed that DNPs hinder the proton translocation through the mitochondrial inner membrane and therefore inhibit oxidative phosphorylation. Their fluorescence quenching properties can help understand and explain their toxicity. Fluorescence quenching of tryptophan was tested using different dinitrophenols such as 2,4-dinitrophenol (2,4-DNP), 4,6-dinitro-orthocresol (DNOC), 2-[(2,4-dinitrophenyl)amino]acetic acid (GlyDNP), 2-(1-methyl-heptyl)-4.6-dinitrophenyl crotonate (Karathan), 2-amino-5-[(1-((carboxymethyl)amino)-3-((2,4-dinitrophenyl)thio)-1-oxopropan-2-yl)amino]-5-oxopentanoic acid (SDN GSH), 2,4-dinitroanisole (2,4-DNA) and 2,4-dinitrobenzoic acid (2,4-DNB). 2,4-DNP and DNOC showed the highest tryptophan fluorescence quenching constant values, these being also the most toxic compounds. The electronic chemical potential value of the most stable complex of 2,4-DNP-with tryptophan is higher than the values of the electronic chemical potentials of complexes corresponding to the derivatives.

The controllable design of multivariate heterojunction with sequential structures is of significant relevance for breaking the performance limit of binary composite photocatalysts. In this work, the novel dual S-scheme ternary-component AgI/Ag 6 Mo 7 O 24 /exfoliated g-C 3 N 4 (ECN) composite was prepared by a two-step in-situ synthetic strategy. The energy band bending at the heterointerface and the formation of dual built-in electric field could be observed due to distinct work functions of different components in the ternary composite. Benefiting from the sequential heterojunction structure, the AgI/Ag 6 Mo 7 O 24 /ECN composite achieved 98.7% removal efficiency of 2-nitrophenol (2-NP) within 70 min under visible light irradiation, and AgI/Ag 6 Mo 7 O 24 /ECN also showed higher degradation efficiency for a variety of organic pollutants such as methylene blue (MB), rhodamine B (RhB), methyl orange (MO), 4-nitrophenol (4-NP), 2-sec-butyl-4,6-dinitrophenol (DNBP) and tetracycline (TC). Notably, •OH and •O 2 - played dominant roles in the AgI/Ag 6 Mo 7 O 24 /ECN set up, which was consistent with the dual S-scheme charge transfer mechanism. In-depth insights for the photodegradation of 2-NP were presented based on a combined DFT study and GC-MS analysis. Additionally, the photoreduction of Ag + in AgI/Ag 6 Mo 7 O 24 /ECN was also evaded by the fast transfer of photogenerated electrons through the dual S-scheme pathway, achieving the effect of killing two birds with one stone., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

Numerous hazardous chemicals from various industrial sources enter the environment daily; some of them can be used both as drugs and as toxins. Many of these compounds, including dinitrophenols, such as 2,4-dinitrophenol (2,4-DNP), 4,6-dinitrocresol (DNOC), and 2-sec-butyl-4,6-dinitrophenol (dinoseb), are widely used pesticides that persist in some contaminated soils. They have been found in groundwater, causing health and environmental hazards, and are subjects of forensic toxicology. Dinitrophenols have multiple biological and noxious effects based on a mechanism of action characteristic to metabolic inhibitors. Although banned, they have pharmaceutical activity and can be purchased on the Internet as ingredients of weight-loss pills. Many death cases have been reported as accidents in agriculture or as overdoses in weight-loss diets. In this article, we discuss legal aspects of dinitrophenol usage, as well as their biological effect and possible mechanisms of toxicity. [ABSTRACT FROM PUBLISHER]

The Aim of the Study: Was to conduct the analysis of patient's clinical observation with acute dinitrophenol poisoning, admitted to a toxicological department of CCH №6 of Izhevsk, Udmurt Republic in 2021 yr. In this clinical case report, a 19 years old girl, who took 20 tablets of dinitrophenol, illegally obtained in online-shop, died. The fatal outcome was realized by the uncoupling of oxidative phosphorylation mechanism and cellular respiration, which in its turn led to serious dystrophic changes in all organs and tissues. Disorders of hemodynamics and blood rheological properties dominated in poisoning pathogenesis, led to congestion, stasis in microcirculatory vessels, hyperpermeability with multiple perivascular hemorrhages in organs, occurrence of piecemeal necrosis in kidneys and liver, nephrosis and nonspecific reactive hepatitis. Production ATP from ADP becomes impossible in these conditions, and respiratory energy chain completely disappears as heat, that explains the heat-increasing and fat-burning effects of dinitrophenol.

Aerobic biodegradation of 2,4-dinitrophenol (2,4-DNP) and 2,6-DNP by an immobilized mixed microbial community in packed-bed reactors was compared for individual components and in a mixture. The reactors efficiently (>97 %) removed higher 2,4-DNP concentrations (up to 43 g m) compared to 2,6-DNP (<80 % only, even when below 5 g m), the former being metabolized with higher degradation rates. Similar loadings of both DNPs were efficiently degraded (>95 %) when treated as a mixture; i.e., the 2,6-DNP removal efficiency was positively affected by the presence of 2,4-DNP when low concentrations of both pollutants were treated. However, an increase of the total DNP concentration to 41 g m led to a sizable drop of the degradation rate in the mixture but not when the substrates were treated separately. [ABSTRACT FROM AUTHOR]

Herein, an ultra-sonication technique followed by a photoreduction technique was implemented to prepare silver nanoparticle-decorated Chitosan/SrSnO 3 nanocomposites (Ag-decorated Chitosan/SrSnO 3 NCs), and they were successively used as electron-sensing substrates coated on a glassy carbon electrode (GCE) for the development of a 2,6-dinitrophenol (2,6-DNP) efficient electrochemical sensor. The synthesized NCs were characterized in terms of morphology, surface composition, and optical properties using FESEM, TEM, HRTEM, BET, XRD, XPS, FTIR, and UV-vis analysis. Ag-decorated Chitosan/SrSnO 3 NC/GCE fabricated with the conducting binder (PEDOT:PSS) was found to analyze 2,6-DNP in a wide detection range (LDR) of 1.5~13.5 µM by applying the differential pulse voltammetry (DPV) approach. The 2,6-DNP sensor parameters, such as sensitivity (54.032 µA µM -1 cm -2 ), limit of detection (LOD; 0.18 ± 0.01 µM), limit of quantification (LOQ; 0.545 µM) reproducibility, and response time, were found excellent and good results. Additionally, various environmental samples were analyzed and obtained reliable analytical results. Thus, it is the simplest way to develop a sensor probe with newly developed nanocomposite materials for analyzing the carcinogenic contaminants from the environmental effluents by electrochemical approach for the safety of environmental and healthcare fields in a broad scale.

Diabetic nephropathy is strongly associated with both increased oxidative stress and kidney tissue hypoxia. The increased oxidative stress causes increased kidney oxygen consumption resulting in kidney tissue hypoxia. To date, it has been difficult to determine the role of kidney hypoxia, per se, for the development of nephropathy. We tested the hypothesis that kidney hypoxia, without confounding factors such as hyperglycemia or elevated oxidative stress, results in nephropathy. To induce kidney hypoxia, dinitrophenol (30 mg per day per kg bodyweight by gavage), a mitochondrial uncoupler that increases oxygen consumption and causes kidney hypoxia, was administered for 30 consecutive days to rats. Thereafter, glomerular filtration rate, renal blood flow, kidney oxygen consumption, kidney oxygen tension, kidney concentrations of glucose and glycogen, markers of oxidative stress, urinary protein excretion, and histological findings were determined and compared with vehicle-treated controls. Dinitrophenol did not affect arterial blood pressure, renal blood flow, glomerular filtration rate, blood glucose, or markers of oxidative stress but increased kidney oxygen consumption, and reduced cortical and medullary concentrations of glucose and glycogen, and resulted in intrarenal tissue hypoxia. Furthermore, dinitrophenol treatment increased urinary protein excretion, kidney vimentin expression, and infiltration of inflammatory cells. In conclusion, increased mitochondrial oxygen consumption results in kidney hypoxia and subsequent nephropathy. Importantly, these results demonstrate that kidney tissue hypoxia, per se, without confounding hyperglycemia or oxidative stress, may be sufficient to initiate the development of nephropathy and therefore demonstrate a new interventional target for treating kidney disease. [ABSTRACT FROM AUTHOR]

Although they are widely used as insecticides, acaricides and fungicides in the agriculture or as raw materials in the dye industry, dinitrophenols (DNPs) are extremely noxious, death cases having been registered. These compounds produce cataracts, lower leucocyte levels, disturb the general metabolism and can cause cancer. It is also assumed that DNPs hinder the proton translocation through the mitochondrial inner membrane and therefore inhibit oxidative phosphorylation. Their fluorescence quenching properties can help understand and explain their toxicity. Fluorescence quenching of tryptophan was tested using different dinitrophenols such as 2,4-dinitrophenol (2,4-DNP), 4,6-dinitroorthocresol (DNOC), 2-[(2,4-dinitrophenyl)amino]acetic acid (GlyDNP), 2-(1-methylheptyl)- 4.6-dinitrophenyl crotonate (Karathan), 2-amino-5-[(1-((carboxymethyl)amino)-3- ((2,4-dinitrophenyl)thio)-1-oxopropan-2-yl)amino]-5-oxopentanoic acid (SDN GSH), 2,4-dinitroanisole (2,4-DNA) and 2,4-dinitrobenzoic acid (2,4-DNB). 2,4-DNP and DNOC showed the highest tryptophan fluorescence quenching constant values, these being also the most toxic compounds. The electronic chemical potential value of the most stable complex of 2,4-DNP-with tryptophan is higher than the values of the electronic chemical potentials of complexes corresponding to the derivatives. [ABSTRACT FROM AUTHOR]

In order to enhance the photocatalytic performance and stability, the various proportions of the size controlled cerium oxide (CeO 2) nanoparticles were dispersed at the pre-synthesized ZnO. Although, the expected dual absorption onsets, probably due to the diminutive difference between the bandgaps of CeO 2 (∼2.9 eV) and ZnO (∼3.1 eV), were not observed however, a blue shift in the bandgap energy of ZnO was witnessed with the increasing surface density of CeO 2 particles. The delayed excitons recombination process with the increasing concentration of CeO 2 nanoparticles was verified by the PL spectra. The structural investigation by Raman and XRD analysis revealed the surface attachment of CeO 2 particles without altering the rock-salt lattice of ZnO. The morphological and fine microstructural analysis established the uniform distribution of evenly sized CeO 2 particles at the surface of ZnO with the discrete fringe patterns of both the entities whereas the XPS analysis confirmed the majority of Ce4+ in dispersed CeO 2. In comparison to pure ZnO, cyclic voltammetric (CV) analysis, under illumination, exposed the supportive role of surface residing CeO 2 particles in eradicating the photo-corrosion of ZnO whereas the chronopotentiometry (CP) predicted the prolonged life-span of the excitons. Compared to pure ZnO, an appreciably high activity was revealed for 10% CeO 2 loading as compared to pure ZnO for the removal of mono and di-nitrophenol derivatives and their mixtures under natural sunlight exposure. The variations in the removal rates in the mixture as compared to individual nitrophenol exposed the structure-based priority of ROS for the respective phenol. The significantly enhanced photocatalytic activity of the composite catalysts revealed the incremental role of surface-mounted CeO 2 entities in boosting the generation of ROS under sunlight irradiation. The experimental observations were correlated and compiled to establish the mechanism of the removal process. [Display omitted] • The controlled size CeO 2 nanoparticles dispersed at the surface of pre-synthesized ZnO. • The surface CeO 2 entities served as the major charge trapping centers for the excited electrons. • The augmented activity highlighted the role of CeO 2 in the intra-band transfer of excitons. • Degradation studies performed with mixture of MNP, DNP and MCP isomers. • The polarity and the intermolecular interactions determine the ease of degradation. [ABSTRACT FROM AUTHOR]

Abstract: The present review paper summarizes the data available in the literature concerning dinitrophenolic compounds and evaluates male reproductive toxicity in experimental animals. Gavage and feeding doses of 2-sec-butyl-4,6-dinitrophenol (dinoseb; CAS No. 88-85-7) manifested testicular toxicity, and 4,6-dinitro-o-cresol (DNOC; CAS No. 534-52-1) showed similar but weaker testicular toxicity in laboratory animals. Consecutive doses of dinoseb and DNOC by gavage seemed to induce spermatotoxicity by disturbing spermiogenesis or the maturation process of sperm in the epididymis, and the most probable target cells of spermatotoxicity were thought to be testicular spermatids in rats. Prolonged exposure to dinoseb and DNOC in the diet also induced testicular toxicity in rats. However, the feeding dose of dinoseb irreversibly affected the early stage of spermatogenesis and produced infertility in rats. On the other hand, 2,4-dinitrophenol (DNP; CAS No. 51-28-5) did not show testicular toxicity in laboratory animals according to available literature. Further studies in laboratory animals with nitrophenolic compounds are required for clarification of their testicular toxicity and for risk assessment in humans. [Copyright &y& Elsevier]

N2O4 was easily impregnated on polyethyleneglycol to give a stable reagent. The polyethyleneglycol-N2O4 (PEG-N2O4) system was used as an effective nitrating agent for the nitration of phenols. Mono- and dinitrophenols can be obtained via direct nitration of phenols in the presence of PEG-N2O4 at room temperature in moderate to high yields. [ABSTRACT FROM AUTHOR]

Abstract: An easy and rapid hollow-fibre supported liquid membrane method (HFSLM) has been developed to extract and determinate the total concentration of four dinitrophenols in environmental water at ngL−1 level. This extraction method provides a high selectivity, short extraction time and very low cost for real samples. It is a three-phase system, aqueous–organic–aqueous, where the organic solvent is held into the fibre pores, being in contact with the two other phases. The organic phase is formed by two different organic solvents, with two different polarities, n-undecane and toluene (1:1). The optimization step was performed using a three-variable Doehler design, involving three factors, stirring speed, fibre length and sample volume. The organic phase composition, as well as the pH of the acceptor and donor phases was also optimized. The extraction equilibrium was reached after 30min, after which essentially the total amount (90–80%) of the four dinitrophenolic compounds were extracted from the sample. Better repeatability and reproducibility at the expense of lower enrichment factors was obtained compared with other methods, employing incomplete extraction during a fixed time. The matrix effect was tested by performing extractions from leachate water and river water. This method is linear in the range 0.1–100μgL−1 in different matrices, with detection limit around 100ngL−1, after extraction of 6mL of sample and using high performance liquid chromatography for final analysis. [Copyright &y& Elsevier]

Abstract: A hollow fiber supported liquid membrane extraction method for the liquid chromatographic determination of dinitrophenolic compounds at ppt levels has been developed. Different variables affecting the extraction process, such as extraction time, shaking speed, acceptor pH, acceptor buffer concentration, salt content and humic acids have been studied. Enrichment factors up to 7000 times were obtained. Validation of the method included calibration experiments and studies of the linearity of the responses in different matrices. Good linearity was obtained in the environmental matrices evaluated. Detection limits range from 6.0 to 8.0ng/L, and the relative standard deviations do not exceed 7% in terms of repeatability. [Copyright &y& Elsevier]

Abstract: A simple and rapid in vitro toxicological assay, utilizing submitochondrial particles (SMP), has been used to evaluate the toxic effects of fifteen herbicides belonging to the phenol and phenoxyalkanoic acid chemical classes. The SMP assay allows the quantitative evaluation of the toxicity of compounds with different mechanisms of action: uncouplers, inhibitors of the enzyme complexes involved in reverse electron transfer and in oxidative phosphorylation and chemicals that alter the membrane structure. The two groups of herbicides showed different levels of toxicity. For phenol derivatives, EC50 values ranged from 0.16μM (ioxynil) to 6.7μM (2,4-dinitrophenol), whereas for phenoxy herbicides EC50 values ranged from 21μM (2,4,5-trichlorophenoxyacetic acid, 2,4,5-T) to 110μM (4-chloro-2-methylphenoxyacetic acid, MCPA). On the average, the toxicity of phenolic compounds is greater than that of phenoxyalkanoic acids by two orders of magnitude. Quantitative structure–activity relationships (QSAR) were developed between EC50 values and various molecular descriptors. The results suggest the existence of different mechanisms of action for the two classes of compounds. The findings obtained for phenolic herbicides are consistent with a protonophoric uncoupling mechanism, whereas for phenoxy herbicides a non-specific mode of action at membrane level can be hypothesized. [Copyright &y& Elsevier]

Natural montmorillonite clay and anthropogenic organic pollutants frequently coexist in the estuarine environment where freshwater from rivers mixes with saltwater from the ocean. In this environment, the sharply changed aqueous chemistry especially salt content could significantly alter the photochemical behaviors of pollutants. However, this process was rarely investigated. In this study, the photodegradation of a representative anthropogenic weight-loss compound 2,4-dinitrophenol in the presence of Fe 3+ -montmorillonite and different halide salts was systematically investigated. Results show that 2,4-dinitrophenol was resistant to photodegradation by Fe 3+ -montmorillonite alone, but the presence of NaCl, NaBr, and sea salts in the system can evoke significant 2,4-dinitrophenol degradation. The enhancement effect was further elucidated as the replacement reaction between the clay associated Fe 3+ and Na  +  which leads to the release of more interlayer Fe 3+ from montmorillonite, resulting in increased production of high active hydroxyl radicals (˙OH) that can substantially damage 2,4-dinitrophenol molecule. In addition, halogen radicals from the reaction of halide ions with ˙OH were also confirmed to participate in 2,4-dinitrophenol degradation. Overall, this study implied that the changed salty condition in the estuarine water could induce the rapid transformation of organic pollutants that move from freshwater and have relatively stable photochemical properties., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

A hybrid Fv fragment of the dinitrophenyl-binding immunoglobulin A (IgA), MPOC315, has been generated by reconstituting a recombinant variable light chain (VL) produced in Escherichia coli with a variable heavy chain (VH) derived from the antibody. The Tyr34 residue of VL was substituted by His in order to introduce a catalytic imidazole into the combining site for the ester hydrolysis. The His mutant Fv accelerated the hydrolysis of the 7-hydroxycoumarin ester of 5-(2,4-dinitrophenyl)-aminopentanoic acid 90,000-fold compared to the reaction with 4-methyl imidazole at pH 6.8 and had an initial rate that was 45 times as great as that for the wild-type Fv. The hydrolyses of aminopropanoic and aminohexanoic homologs were not significantly accelerated. Thus a single deliberate amino acid change can introduce significant catalytic activity into an antibody-combining site, and chemical modification data can be used to locate potential sites for the introduction of catalytic residues.

Glucocorticoids induce an alteration of the surface of hepatoma tissue culture (HTC) cells as expressed by changes in cell electrophoretic, antigenic, and adhesive properties. The alteration is assayed by the increased adhesiveness of induced cells for a glass surface. The induction process has a lag period of about 3 hr and attains a plateau level after 24-30 hr when 50-80% of the steroid-treated cells are firmly adhered. Less than 10% of untreated cells adhere under the same conditions. Induction is inhibited by actinomycin D and cycloheximide, demonstrates both pH and temperature dependence, and responds to changes in steroid concentration and structure. By contrast, the attachment per se of preinduced cells is not affected by inhibitors of RNA and protein synthesis, fluctuations of temperature and pH, and the presence or absence of the hormone. When the induction process is reversed by removal of steroid or addition of actinomycin D, preinduced adhesiveness is lost with a half-life of 13-24 hr, but in the presence of cycloheximide the loss is accelerated (t(1/2) 3-5.5 hr). These results suggest that glucocorticoids induce the biosynthesis of a protein which either modifies the cell surface (an enzyme) or is incorporated into surface structures (structural protein).

These in vitro studies were conducted in an attempt to elucidate the mechanism of how cell-free supernatant fluids obtained from PHA-stimulated human lymphocytes cause destruction of cells. The undiluted supernatant fluids with high activity exerted a nonspecific cytotoxic effect on many different continuous cell lines. However, upon dilution, a wide spectrum of cell sensitivities was observed. These studies suggest human lymphotoxin acts by first absorbing to receptors on the target cell plasma membrane. The next effect is shut-down of cellular DNA synthesis, followed later by a decrease in cell numbers and finally, cellular destruction. Once sufficient LT has bound to the target cell surface, the cytopathic effect is irreversible. A role for LT in lymphocyte-mediated tissue destruction is discussed. © 1973.

Mast cells (MCs) are an important treatment target for high-affinity IgE Fc receptor (FcεRI)-mediated allergic diseases. The plant-derived molecule 4-methylumbelliferone (4-MU) has beneficial effects in animal models of inflammation and autoimmunity diseases. The aim of this study was to examine 4-MU effects on MC activation and probe the underlying molecular mechanism(s). We sensitized rat basophilic leukemia cells (RBLs) and mouse bone marrow-derived mast cells (BMMCs) with anti-dinitrophenol (DNP) immunoglobulin (Ig)E antibodies, stimulated them with exposure to DNP-human serum albumin (HSA), and then treated stimulated cells with 4-MU. Signaling-protein expression was determined by immunoblotting. In vivo allergic responses were examined in IgE-mediated passive cutaneous anaphylaxis (PCA) and ovalbumin (OVA)-induced active systemic anaphylaxis (ASA) mouse models. 4-MU inhibited β-hexosaminidase activity and histamine release dose-dependently in FcεRI-activated RBLs and BMMCs. Additionally, 4-MU reduced cytomorphological elongation and F-actin reorganization while down-regulating IgE/Ag-induced phosphorylation of SYK, NF-κB p65, ERK1/2, p38, and JNK. Moreover, 4-MU attenuated the PCA allergic reaction (i.e., less ear thickening and dye extravasation). Similarly, we found that 4-MU decreased body temperature, serum histamine, and IL4 secretion in OVA-challenged ASA model mice. In conclusion, 4-MU had a suppressing effect on MC activation both in vitro and in vivo and thus may represent a new strategy for treating IgE-mediated allergic conditions.